Chemistry of Carbohydrates INTRODUCTION Carbohydrates are the most abundant organic molecules present in nature, They make up majority of the organic matter on earth, because of their multi- ple roles in all forms of life. The empirical formula of the carbohydrates is (CHO), hence the name ‘hydrates of carbon’, although it is a misnomer, be- cause there are certain noncarbohydrates (e.g. formaldehyde — CHyO) having this formula, while many carbohydrates (e.g. deoxyribose — CsHy)0,) do not follow this formula. Carbohydrates include compounds with relatively small molecules like simple sugars (monosaccharides and disaccharides) as well as macromolecular sub- stances such as starch, glycogen and celluloses. Whenever the word ‘sugar’ is mentioned, the idea of energy conjures up in our minds. On learning the structure-function relationship about biomolecules, one comes to know that each and every molecule differs structurally and hence functionally. Table sugar is sucrose, whereas the sugar predominantly present in fruits is fructose. Small changes in structural configuration lead to changes in the functional properties. Thus, when there is an (alpha) a-linkage between glucose molecules, the result- ing homopolysaccharide is starch/glycogen, which is a utilisable storage form of energy in plants and animals. When this linkage between glucose molecules is B (beta), the molecule formed is cellulose, which is not digestible in the human body. Furthermore, only the B-form of glucose can be absorbed by the body, whereas the o-form remains unabsorbed. Carbohydrates not only serve as major sources of energy but also function as precursors for the synthesis of lipids, amino acids, glycoproteins and proteogly- cans in the body. In biochemistry, the functions and behaviour of molecules are highly dependent on the structure of the molecule. Carbohydrates have a wide range of functions, which may be summarised as follows: * They serve as a principal energy fuel for the cell, e.g. glucose. ° They serve as a storage form of energy, e.g. glycogen. * Certain metabolic intermediates are carbohydrates, e.g. glyceraldehyde. * ATP, the university energy currency, is a phosphorylated sugar derivative, as are many coenzymes. * Some carbohydrates serve as genetic material, e.g. ribose and deoxyribose sugars form part of the structural framework of RNA and DNA. * Many polysaccharides serve as structural components of bacterial and plant cell walls and are also part of the exoskeleton of arthropods. WWical Biochemistry: Preparatory Manual for Undergraduates i inked to many proteins and lipids, and they play re also linked y pre lipids + Garbohydiattrole in the stricture and function of the biological membranes, an imporuzell recognition and intercellular communication. eg. in cel E : ecules, which perform different functions s include a host of molecules, learn to aoe that follow will help you. nd swers Q.1. Define and classify carbohydrates. Ans. + Carbohydrates are defined as polyhydroxy derivatives of aldehydes or ketones. * Carbohydrates are classified into four groups: Carbohydrates | Monosaccharides Disaccharides Oligosaccharides __ Polysaccharides * Monosaccharides (simple sugars) * Monosaccharides are those carbohydrates that cannot be further hydro- lysed. These are similar to amino acids present in proteins. * As stated earlier, the general formula of carbohydrates is Cyl1s,0,, Depend: ing upon the number of carbon atoms they possess, they are subdivided! into tnioses (30), tetroses (4C), pentoses (5C), hexoses (6C) or heptoses (7C). * Also, depending upon whether an aldchyde or a ketone group is present, they are divided as aldoses or ketoses (Table 3.1). Aldoses Ketoses. Tiiose C3H05 Glyceraldehyde Dihydroxyacetone Tetrose CHO. Erythrose Erythrulose: Pentose CsH1005, Ribose Ribulose Hexose CoH 20s, Glucose. Fructose ‘Sedoheptose Sedoheptulose * Disaccharides © The linkage between two monosaccharides is called a * Disaccharides are those sugars that y ghcosidic linkage. } field two molecules of the same or differ- ent monosaccharides when hydrolysed, * General formula is C,H2,0,y. Disaccharides Sucrose Lactose MaltoseChapter 3 — Chemistry of Carbo © Examples Sucrose: glucose + fructose (nonreducing sugar) Lactose: galactose + glucose (reducing sugar) Maltose: glucose + glucose (reducing sugar) © Oligosaccharides © These are the carbohydrates that yield 2-10 monosaccharide units upon hydrolysis, e.g. maltotriose. © They serve as parts of blood group antigens. These are not discussed in detail as they account for a very small percentage of naturally occurring carbohydrates. * Polysaccharides © Polysaccharides yield more than 10 molecules of monosaccharide units upon hydrolysis. General formula is (CsH90;),,. Polysaccharides are further divided into the following types: Polysaccharides Homopolysaccharides Heteropolysaccharides (i) Homopolysaccharides: These are the polymers of the same monosaccha- ride units, e.g. starch, glycogen, inulin, cellulose, dextrin, dextran. (ii) Heteropolysaccharides: These are the polymers of different monosaccha- ride units or their derivatives, e.g. mucopolysaccharides. Q.2. Write a note on monosaccharides. Explain their sources and biochemi- cal importance, Ans. Monosaccharides (or simple sugars) are water-soluble, insoluble in lipid solvents, odourless crystals with sweet taste. Sugars are mainly composed of hydroxyl and carbonyl groups. The source, biochemical importance and physi- ological reactions of monosaccharides are summarised in Table 3.2. Significance Importance o-Glucose Fruits Principal form of Present in urine (glycos- | ‘energy in the body uria) in patients with diabetes mellitus due to raised blood glucose | | (hyperglycaemia) | orwaseieveaas honey Is converted to Hereditary fructose | glucose in liver and intolerance leads to intestine fructose accumulation and hypoglycaemia Failure to metabolise ‘Sweeter than glucose 0-Galactose Is converted to Hydrolysis of lactose glucose galactose leads to Synthesised in galactosaemia and mammary glands to cataract make lactose Continued1A Medical Biochemistry: Preparatory Manual for Undergraduates ‘Sugars Sources. Significance Importance p-Mannose Hydrolysis of ‘Sugar occurring in Used to treat urinary mannans and gums glycoproteins tract infections D-Ribose Nucleic acids Structural element of _Excreted in pentosuria nucleic acids and co-enzymes D-Ribulose Formed in body Intermediate in hexose monophosphate shunt p-Arabinose Cherry gums, gum —_—_Energy and carbon. arabic source in bacteria used as an additive in food Inhibits sucrase D-Xylose Wood gums, Part of glycoproteins Found in urine in proteoglycans essential pentosuria Q8. Write a note on stereoisomerism. Give suitable examples. Ans. © Compounds that have the same structural formula but differ in spatial configuration are known as stereoisomers. * A carbon atom that is bonded to four different chemical groups or atoms is known as an asymmetric carbon. * All monosaccharides contain at least one asymmetric carbon. In glyceraldehyde, the second carbon is the asymmetric carbon atom. © The presence of an asymmetric carbon atom allows the formation of isomers. The number of possible isomers of a compound depends on the number of ; asymmetric carbon atoms (n); it is equal to 2". * Glucose has four asymmetric carbon atoms. Therefore, it can form 2! = ‘ 4 q G isomers. Glyceraldehyde has one asymmetric carbon atom and forms two isomers. The important isomers are as follows: (i) Epimers * Stereoisomers differing in configuration of the OH group on only one asymmetric carbon are known as epimers; e.g. mannose and galactose are epimers of glucose. Mannose and glucose are C-2 epimers. © Their structure differs in the position of the ~OH group only at the car- q 5 4 bon atom at the second position. = © Galactose and glucose are C4 epimers (Fig. 3.1). Note, Mannose and galactose are not epimers of each other as they differ in the position of OH group at two positions. (ii) Enantiomers © These are the stereoisomers that are mirror images of each other They are also called p- and t-isomers, e.g. p-glyceraldehyde and tglyceraldehyde are enantiomers (Fig. 3.2). * When the -OH group on the penultimate carbon atom is on the right, it is said to belong to the p-series. p-glucose belongs to this series. The majority of the sugars in humans are p-sugars. in glycosaminoglycans (GAGs). Glucose found in humans is always peglucose, =a + Exceptions are L-fucose found in glycoproteins and t-iduronic acid foundCHO CHO CHO | H- 0-H H-¢- OH HO} ¢- H Of Ho— 0-H HO=¢—H HO]- ¢- H H-0— OH H—C— OH namo Bisa H-G—OH CH,OH CH,OH CH,OH D-Galactose D-Glucose D-Mannose C4 C2 Fig. 3.1. Epimers. 9 9 I {I Hee Hee Hal BCH CH2OH CH20H D-Glyceraldehyde Asymmetric carbon atom t-Glyceraldehyde Fig. 3.2 Enantiomers. (iii) Optical isomers (D- and L-isomers) © The presence of asymmetric ca the compound. * When a polarised light is passed through a solution exhibiting optical activity, the plane of the light is rotated to the right or left. © When it is rotated to the right, the compound is called dextrorotatory (denoted as dor +), and when rotated to the left, the compound is called laevorotatory (denoted as or -). © When equal amounts of dextro- and laevorotatory isomers are present, the resulting mixture has no optical activity and is called a racemic mixture. Mutarotation * A freshly prepared solution of monosaccharide shows a gradual change in its optical activity until it becomes stable on standing. This phenome- non is called mutarotation. ° The changes are due to the presence of anomeric forms of sugar in solution. For example, in an aqueous solution of glucose crystallised from cold ethanol at room temperature, a-D-glucose separates, having an tial specific rotation of +112°, which changes to a fixed rotation of +52.5°. * On the other hand, when crystallisation takes place from water at a temperature above 98°C, a B-p glucose separates and the initial specific rotation of +19° changes to a fixed rotation of +52.5° (Fig. 3.3). © This occurs due to the opening and closing of the hemiacetal ring in the solution. When the ring opens, a straight-chain aldehyde is formed. On losing water, the anomeric carbon may have a hydroxyl group in either the a or the B-position. The a- or B-form slowly achieves an equilibrium, resulting in a fixed rotation (+52.5° in the above example). At equilibrium, 64% of B-glucopyranose and 36% of a-glucopyranose are produced. bon atoms also confers optical activity onMedical Biochemistry: Preparatory Manual for Undergraduates. H [OH Z gro eel Bagel : —> ale —> > HO-—C—-H HO: H Saree H-C—-OH Mat H-C—OH CHZOH CH,OH, -0.Glucose (0): a-(0)-Glucose que Ae B-(0)-Glucose rc (1° Fig. 3.3 Mutarotation. (v)_Hemiacetal formation © Under acidic conditions, aldehydes combine with one or two of the hydroxyl groups of an alcohol, forming a hemiacetal or an acetal, respectively. i H Hy H \ I R—C=0 +ROH—> R=C-OH R—C=0 +2R0H—> R— Goon! ‘ o-F on’ £ A hemiacetal An acetal * The cyclic hemiacetals lead to the formation of a ring structure. (vi) Pyranose and furanose ring structures © In 1929, Haworth proposed a six-member ring form of a sugar called ‘pyranose’ because pyran possesses a ring of five carbons and an oxygen. © Similarly, Haworth called sugars containing a five-member ring ‘furanoses’ because furan contains a ring of four carbons and one oxygen. * Less than 1% of the monosaccharides with five or more carbons exist in the open-chain form. In solution, they exist in the ring forms (i.e. pyranoses and furanoses). Glucose normally exists in a pyranose form, whereas fructose is present in a furanose form (Fig. 3.4). Pyran ring Furan ring OH HOH,C CH,OH H Q [OH on pa H OH Ht 1H -D-Glucopyranose -D- Fructofuranose Fig. 3.4 Ring structures of glucose and fructose.Chapter 3 — Chemistry of Carbohydrates — Anomers © Anomeric carbons are the asymmetric carbons (C-l in case of glucose) that are created by cyclisation at the carbon bound to oxygen in hemiacetal formation. * A ring structure of glucose is formed by interaction of carbonyl group with alcoholic group. This ring formation results in the creation of an anomeric carbon at G-l in aldose and C2 in ketose sugars. (vili) a and B-Anomers © The pyranose form can have -H of interacting -CHO on the left and -OH on the right, in which case it is called -anomer; if -OH is on the left and -H on the right, it is called a B-anomer (Fig. 3.5). * Enzymes can differentiate between these configurations and are specific for only one of these. Thus, cellulose having a B(I>4) linkage cannot be hydrolysed by amylases, which recognise only « (I4) linkages. iv [ou] mi I I H-e=0H H-G-OH HosesH HO-C-H HaaoH H-C-OH | Si — H-C CH,OH CH,OH a-D-Glucopyranose -0-Glucopyranose Fig. 3.5 Anomers of o-glucose. © The aldehyde and ketone groups react with the alcohol group of the same sugar to form hemiacetal and hemiketal rings, respectively. Glucose is predominantly present in the e-form. Q.. Describe the chemical properties of monosaccharides. Give details of the tests performed using these properties. Ans. © Given the importance and the multiple types of monosaccharides, it is neces- sary for us to identify ways to distinguish between them. * A number of chemical properties of monosaccharides have been used to this purpose. Amongst these, the important ones are as follows: (a) Reducing property of monosaccharides (b) Action of strong acids on monosaccharides (c) Oxidation reactions (d) Reduction reactions (e) Osazone formation (O) Glycoside formation (a) Reducing properties ° Sugars that can be oxidised at the carbonyl carbon are known as reducing sugars. © The sugars react with chemical reagents due to the presence of free carbonyl groups. In an alkaline medium, the sugar forms enediols, which reduce Cu*’, Ag*, Fe*® and Bit, and gets oxidised to sugar acidsMedical Biochemistry: Preparatory Manual for Undergraduates © Such sugars are called reducing sugars, e.g. glucose and fructose. The reducing action of sugars in an alkaline medium is used for both qualitative and quantitative determinations of sugars. Some examples are as follow: (i) Barfoed’s test = Barfoed’s reagent contains a solution of copper acetate in acetic acid. = With Barfoed’s reagent, monosaccharides react very fast as compared to disaccharides because of the presence of the weak acid ~ A red precipitate is formed by the monosaccharides but not by disaccharides. = Barfoed’s test helps to distinguish between monosaccharides and disaccharides. (ii) Benedict’s qualitative test = This test is used to detect reducing sugars in urine. — Gynaecologists and endocrinologists advice the performance of Benedict's qualitative test in their ward-side laboratories. This is because it is the simplest way in which gestational diabetes can be detected. Colours: Degree of glucosuria Cone. (g/dl) | Green + 05 Yellow ++ 1.0 i Orange tet 15 1 =2.0 = The reagent contains Cu*? (cupric sulphate), alkali (sodium carbonate) and sodium citrate. = Sodium citrate prevents the precipitation of copper hydroxide ‘or carbonate by forming soluble complexes with Cu?* ions. - On heating with Benedict’s reagent, the reducing sugar forms enediol in alkaline medium, which then reduces Cu** to Cur. ~ This then forms a yellow-coloured precipitate depending on the | concentration of the solution. (iii) Fehling’s test — In Fehling’s test, KOH is used as the alkaline medium and potas- sium tartrate is used instead of sodium citrate. Reducing sugar + Alkali ——> Enediol + Cu” » cu’ « sugar acids OH i | Cuprous oxide 2 cul + cu,0+H,0 ‘Cuprous hydroxide | Red ppt. (b) Action of strong acids on monosaccharides * Sugar when heated with strong mineral acids loses water and forms furfurals.Chapter 3 — Chemistry of Carbohydra ° Furfurals may condense with phenolic compounds such as «-naphthol, thymol and resorcinol to produce coloured complexes. This property forms the basis for the identification of sugars. Examples: (i). Molisch’s test: general test for identification of carbohydrates at the junction of two layers. o-Naphthol Sugar solution + HyS0, ————> Red violet ring (ii) Setiwanoff's test HCl Fructose —————> Cherry red colour Resorcinol Note. This testis used for distinguishing between glucose and fructose. A word of caution: if boiled for longer time, glucose can also give positive results, so be carefull (iii) Bial’s test Bial’s test is used for the detection of pentoses. Fructose gives red colour with Bial’s reagent. HCL, Orcinol Ribose —————> Green colour (blue/brown on standing) FeCl, (c) Oxidation to form sugar acids Aldoses can be oxidised to different compounds depending upon the reactants. Examples are as follows: * p-Glucose is oxidised to an aldonic acid called gluconic acid. Simi- larly, mannose and galactose form mannonic and galactonic acids, respectively, with bromine water. * Strong oxidising agents such as concentrated HNOs oxidise the aldehyde as well as the primary alcoholic group to produce saccharic acid. For ex- ample, glucose on oxidation produces glucosaccharic acid; galactose on oxidation with HNOs produces galactosaccharic acid or mucic acid. © Under specific conditions, when the aldehyde group is protected and the sugar is oxidised, uronic acid is produced. For example, upon oxidation, glucose gives rise to glucuronic acid, while mannose and galactose form mannuronic acid and galacturonic acid, respectively. (COOH, CHO oon | | t i i CH,OH (COOH (COOH Aldonic acid Uronic acid Saccharic acid © Here ‘R’ represents the rest of the carbon atoms. In the body, glucuronic acid is formed in the liver and is used for detoxication. © p-Glucuronic acid occurs as a constituent of certain mucopolysaccharides. It is important in detoxication mechanism, where bilirubin is linked with p-glucuronic acid. (d) Reduction to form sugar alcohols © Aldose monosaccharides may be reduced to their corresponding alco- hols by reducing agents such as sodium-amalgam or by hydrogen under high pressure in the presence of a catalyst.20 Medical Biochemistry: Preparatory Manual for Undergraduates © Aldoses yield the following alcohol IG te ee Ea Sorbitol Na/Hg Hy D-Galactose ———> )-] ‘i lactose <7 Gg_ D-Dulcitol D-Mannose —=_> p, i Na/Hig, PMannitol * Ketoses yield two alcohols because of the formation of a new asymmetric carbon in position C-2. © Therefore, fructose yields a mixture of mannitol and sorbitol. Hi, Fructose ——2 > p-Mannitol + p-Sorbitol Na/Hg Important reactions of this type are as follows: (i) Reduction of glycerose to glycerol (ii) Reduction of ribose to ribitol, which is part of riboflavin (iii) Reduction of glucose to ascorbic acid (vitamin C) ‘Sugar alcohol__Importance Sorbitol '35%-60% sweetness of glucose. Used as a sweetener. Can be accumulated in tissues such as the eye lens, sciatic nerves in case of galactosaemia and diabetes mellitus. Mannitol Found in plant extracts. Has 50% sweetness of sucrose. Administered intravenously as an osmotic diuretic in patients with acute renal failure. Also used to relieve increased pressure in cerebrospinal fluid. Glycerol Component of triacylglycerol and phospholipids. Ribitol (Certain bacteria use this molecule as an energy source. Itis also a Constituent of riboflavin, a component of flavin coenzymes. ‘Twice as sweet as sucrose. May be used in prevention of dental caries. Xylitol (e) Osazone formation (Fig. 3.6) * The presence of free carbonyl (of aldehyde or ketone) group is essential for this reaction. H- GE] NENH= GeHs] H-G-OH R HN=NA=C5 Hy Phenylhydrazine HE] _NENA=O; Hi] H-CE5_NE=NH=G5H,| R Glucosazone Fig. 3.6. Formation of glucosazone with phenylhydrazine.Chapter 3 — Chemistry of Carbohydrates * A monosaccharide or reducing disaccharide is first heated with GF phenylhydrazine in acetic acid to form sugar-phenylhydrazine, which reacts with more reagents to yield the particular _osazone-forming characteristic crystals of specific Sl A ap ° This test helps to distinguish be- tween different sugars. Maltose osazones © Glucose, fructose and mannose Figrat7amcsaconess form the same osazones (i.e. form the crystals of needle shape); malt- ose forms crystals of sunflower shape and lactose forms crystals known as ‘balls of spicules’ (Fig. 3.7). (f) Glycoside formation © Glycosides are formed by interaction between a monosaccharide or a monosaccharide residue and the hydroxyl group of a second com- pound that may or may not be a monosaccharide. © The noncarbohydrate group is called aglycone. The glycosidic bond is an acetal link as it results from a reaction between a hemiacetal and—OH group. Glucose osazones Lactose osazones Fructose osazones: ‘Sugar-H+R-OH ——> Sugar-R (Glycoside) + H2O © If the sugar residue is glucose, the derivative is called glucoside. If it is fructose, it is called fructoside. © Ifthe side chain (R) is another sugar, the glycoside becomes a disaccharide. Sugar;-H + OH- Sugarp ——» Sugar;-Sugara+ HzO If the side chain is a sterol, it forms the basis for the formation of drugs, such as streptomycin, Q5. What are the uses of glycosides? Glycosides are found in many drugs and spices. They are also constituents of animal tissues. Aglycone may be methanol, glycerol, sterol or a phenol. Glycosides are important in medicine because of their action on heart (cardiac glycosides). These include derivatives of digitalis and ouabain. Ouabain is an inhibitor of Na*, K*-ATPase of cell membranes. © Other glycosides include antibiotics such as streptomycin. Phlorhizin is a glycoside that blocks the transport of sugars across the mucosal cells of the small intestine. Q Ans. There are many derivatives formed from monosaccharides, which are biologically important. * When the chemical composition of monosaccharides is altered, we obtain certain other biomolecules which, by themselves, have important biological functions. ¢ The point that differentiates these molecules from those we have studied before is that these derivatives do not adhere to the (C,H»,0,) formula. What are the derivatives of monosaccharides?Medical Biochemistry: Preparatory Manual for Undergraduates (i) Deoxy sugars © Deoxy sugars represent sugars in which the oxygen of -OH group has been removed, leaving the hydrogen. Thus, -CHOH or -CH,OH becomes ~CHy or -CHs. © Deoxy sugars of biological importance are 2-deoxyribose found in nucleic acid (DNA), and 6-deoxygalactose found as a constituent of bacterial polysaccharides. (i) Amino sugars ‘© Sugars in which hydroxyl group is replaced by an -NHp group are called amino sugars, e.g. glucosamine and galactosamine. * Amino sugars occur as N-acetyl derivatives in a number of important biological substances. © N-acetyl derivative of p-glucosamine occurs as a constituent of hyaluronic acid, heparin and some blood group substances. © Galactosamine occurs as N-acetylgalactosamine in chondroitin sulphate. Cer- tain antibiotics such as erythromycin and carbomycin contain amino sugars. (iii) Esters of sugars © Esterification of carbohydrates with phosphoric acid isa common reaction in body metabolism. © Glucose 6-phosphate and glucose I-phosphate are good examples. ATP donates the phosphate moiety for ester formation. (iv) Sugar acids © These have been described earlier in this chapter. a (¥) Glycosides * Glycosides have been discussed earlier in this chapter. Q7. What are disaccharides? Give details of the three common disaccharides of biological importance. Ans. When two monosaccharides are combined together by a glycosidic linkage, a disaccharide is formed. Three most common disaccharides of biological importance are as follows: (a) Maltose (b) Lactose (c) Sucrose (a) Maltose(malt sugar) ° The malt present in ‘Bournvita’, ‘Boost’, etc., is maltose. Maltose is formed when starch is hydrolysed by a-amylase. * It consists of two molecules of glucose bonded by an «(I->4) glycosidic linkage (Fig. 3.8). CHO OH Free -OH group fy e0-Glucose 4), Fig. 3.8, Maltose (O-a-p-glucopyranosyl-4-c-b-glucopyranoside). DEChapter 3 — Chemistry of Carbohydrates CH.OH H H OH H 2 [OH] Free -OH group H OH B-0-Galactose a-D-Glucose Fig. 3.9 Lactose (1--b-galactopyranosyl-a-4-p-glucopyranoside). © It forms characteristic ‘sunflower’-shaped crystals of osazones with phenylhydrazine. ° The anomeric carbon of one glucose being free, maltose demonstrates all the properties of a reducing sugar. Isomaltose is also a reducing disaccharide. It consists of two glucose units. © The linkage is a(1-¥6) glycosidic. It is formed by the partial hydrolysis of glycogen and starch. (b) Lactose (milk sugar) ® Lactose is found in milk and therefore is called milk sugar. It is synthesised by mammary glands. ® It consists of galactose and glucose molecules linked by a B (14) glycosidic linkage (Fig. 3.9). The linkage is between C-l of B-galactose and C-4 of a-glucose. The anomeric carbon of glucose is free. Therefore, lactose is a reducing disaccharide, © It forms osazones of characteristic shape referred to as ‘ball of spicules’. It gives positive reducing tests. * As lactose contains galactose as one of the components, it yields mucic acid on treating with concentrated HNOs. (c) Sucrose (cane sugar, table sugar, saccharose and beet sugar) © Sucrose is widely distributed in plants, particularly in sugarcane and sugar beet. The sugar used on the table is sucrose. Chemically, it is made up of one unit of glucose and one unit of fructose. As both groups, i.e. aldehyde group of glucose and ketonic group of fructose, are involved in the formation of a glycosidic linkage, there is no free reducing group. The linkage is a, (1-2) glycosidic, between C-1 of o-glucose and C2 of B-fructose. Hence, sucrose is a nonreducing sugar (Fig. 3.10). oonon yy % cH,OH cH,OH = a A) 4 H O 4 “ H as RY 1 be yor™ OH H ° ae CH,OH H OH OH H -D-Gl Sia Sucrose B-D-Fructose Fig. 3.10 Sucrose (1-a-D-glucopyranosyl-2--p-fructofuranoside).Medical Biochemistry: Preparatory Manval for Undergraduates k ry (-92°) whereas that of glucose is dextrorotatory (+52.5°), + The resultant product of hydrolysis shows * This change from dextrorotation to lae sucrose is called ‘invert sugar’, * the net results laevorotatory resulting in change of rotation from positive to negative. laevorotation. ‘vorotation is called inversion. Hence, Q9. Comment on the degree of sweetness of the different sugars. Ans. Sucrose is considered to be 100% Sweet, and relative to this, lactose 20%, Blucose is 50% 70%, fructose is 130%-170% — the reason why fruits taste sweet, Sorbitol is 40%-50%. Saccharine, the artificial sweetener, has a sweetness of 40,000. Q.10. What are polysaccharides? Wri including their clinical uses, occurr: Ans: Polysaccharides are high-molecularweight compounds that can be divided into the following two types: { Homopolysaccharides (homoglycans) * Heteropolysaccharides (heteroglycans) The de tails of the homopolysaccharides are as follows (Table 3.3) Starch Substance Amylose Dextrin _Dextrans Branched Chainof Branched D-glucose p-glucose and un- units after units branched Obtained chain of every 8-10 Itisob- from bacteria glucose afterevery glucose tained from Molecules 25-30 Molecules hydrolysis glucose of starch molecules. Large amount of (156) branching Structure _Helically coiled un- branched Polymer of o-glucose Branching a(14) (154) a (14) Ba) (1-36) a(154) a (136) a(196) a (144) a (13) Colour with Deep blue Violet Deepred No-colour Erythrodex- No colour iodine trins ~red colour Achroodex- trins ~no colour(a) (c) Chapter 3 — Chemistry of Carbohydrates (14) (1-6) Amylose ‘Amylopectin Fig. 3.11 Representation of the two components of starch. Starch * It is a polysaccharide that occurs in the plant kingdom. Its important sources are cereals, grains, potatoes, carrots, millets and legumes. It is a storage polysaccharide and isa source of energy. * Starch is a polymer of glucose. It is a mixture of amylose (159%-85%) and amylopectin (65%-85%).. * Amylose consists of a long helically coiled unbranched chain of glucose molecules joined by a(I—4) linkages. Molecular weight ranges from 10,000 to 100,000, * Amylopectin is insoluble in water. Molecular weight ranges from 50,000 to 1000,000. Amylopectin consists of a branched chain of glucose units linked by an ax(I4) linkage, and branching occurs via an a(I-36) linkage between two glucose units of neighbouring straight chains. © The branching points occur at intervals of approximately 25-30 ad-glucose units (Fig. 3.11). * A solution of amylose gives deep blue colour with iodine. Amylopectin gives violet colour with iodine. Glycogen * Itisa storage polysaccharide present in muscle and liver of animals. Itis therefore called animal starch. * Glycogen in the liver is broken down to glucose and passed into the blood stream for use by the tissue. Glycogen in muscle is a good source of energy. * Glycogen is a branched polysaccharide of d-glucose units (Fig. 3.12). Glucose units are joined by «(I—>4) gly- cosidic linkages, and branching occurs at branch points via a(I96) linkages. * Branching occurs at every 8-10 a d-glucose units. Glycogen is not destroyed by hot, strong KOH or NaOH solution, and it is this property that is used in determining it quantita- aan tively in tissues. i r * Glycogen gives deep red colour with G2,Si12, Fenveseniation of iodine. Dextrins * Dextrins occur in tissues of all starch-producing plants. * They also occur in honey and are important constituents of various foods. * They are used exclusively as adhesives and binders. * Dextrins are formed by partial hydrolysis of starch. The hydrolysis can be carried out by acid or the enzyme, amylase. * Dextrins formed from the hydrolysis of amylose have unbranched chains, while those from amylopectin are branched.26 Medical Biochemistry: Preparatory Manual for Undergraduates * They are generally soluble in water and can be precipitated by the action of alcohol and have a sweet taste. © The highly branched dextrins give red colour with iodine: * Depending on the degree of hydrolysis, different colours are obtained with iodine. (d) Cellulose © Gellulose is a chief constituent of plant cells and is the most abundant carbohydrate in nature. It forms the basis of vegetable fibres like cotton and the jute. It is a polysaccharide composed of repeating glucose units, joined together by B(4) linkages. Gellulose is not soluble in water, dilute acids, alkalies and organic solvents. Gellulose cannot be digested by human body. The human digestive tract does not contain the enzyme, cellulose that can attack the B(I4) link: ages in cellulose. Cellulose forms an important part of human diet as it acts as roughage. It is also postulated that cellulose helps prevent colonic cancer, It also has the capacity of conferring a feeling of satiety (feeling of fullness). * Cellulose gives red colour with iodine. (e) Dextrans © Dextran isa complex, branched polysaccharide made up of many glucose molecules joined into chains of varying lengths from 10 to 150 kDa. : * It is synthesised from sucrose by certain lactic acid bacteria, the best if : known being Leuconostoc mesenteroides and Streptococcus mutans. a © The straight chain consists of a(196) glycosidic linkages between glucose molecules, while branches begin from (1-4) linkages [and in some cases, «(1-92) and (1-93) linkages as well]. () Inulin © It occurs in tubers of dahlias, onion and garlic. © Icis a polymer of d-fructose. It can be hydrolysed to d-fructose by acid or by an enzyme inulinase. * It gives no colour with iodine. Inulin has no dietary importance. © Inulin is a homopolysaccharide, which has repeating units of fructose. (g) Agar * Agar occurs in sea weeds. It is the material that confers the gelatinous property to the agar. * Itis made up of repeated units of galactose, which is sulphated. It is used as a laxative to treat constipation. i * Purified agar is used for bacterial cultures. (h) Chitin * Itis present in the exoskeleton of invertebrates such as crabs, lobsters and insects. * Chitin consists of repeating units of N-acetyl-d-glucosamine joined by B(1-+4) glycosidic linkages (Fig. 3.13). * It is similar to cellulose except for the presence of an acetylated amino group in the C2 position. * So, it can be seen that just the addition of acetyl and amino groups alters the composition of substances completely. Q11. What are dietary fibres and how do they affect health? Ans. Dietary fibres are the indigestible portions of plant food that move food through the digestive system, absorbing water and easing defecation. Dietary OEE EeeH CY H H 0, oH | ou if H H H NHCOCH, H NHCOCH, L_____N-Acetylglucosamine N-Acetyiglucosamine =9—J n Fig. 3.13 Chitin. fibre consists of nonstarch polysaccharides such as cellulose and many other plant components such as dextrins, inulin, lignin, waxes, chitins, pectins, beta-glucans and oligosaccharides. The term ‘fibre’ is somewhat of a misnomer, since many of the so-called dietary fibres are not fibres at all. Earlier, the term ‘roughage’ was used. Diet naturally high in fibre can be considered to bring about six main physi- ological changes: © Improvement in gastrointestinal health © Improvement in glucose tolerance and the insulin response Reduction of hyperlipidaemia, hypertension and other coronary heart disease risk factors Reduction in risk of developing some cancers Increased satiety and hence some degree of weight management Insufficient fibre in the diet can cause constipation and possibly lead to the development of haemorthoids or anal fissures Q.12, What are the clinical applications of dextran? Ans. Dextran in intravenous solution provides an osmotically neutral fluid. In the body, itis digested by cells into glucose and free water. It is occasionally used to replace lost blood in emergency situations, when replacement blood is not available. It must be used with caution as it does not provide necessary electro- lytes and can cause hyponatraemia or other electrolyte disturbances. Q.13. What is inulin and what are its uses? Ans. Inulin is a polyfructosan (polymer of fructose) and it is used for the deter- mination of glomerular filtration rate (GFR). The inulin clearance test is a renal function test. It is also used to measure body water volume. Q.14. What are heteropolysaccharides? Briefly explain. Ans. * Under this category, we will study the following three types: (i) Glycoproteins ) Proteoglycan (iii) Glycolipids * The difference between glycoproteins and proteoglycans lies in which group is the majority partner. © When the protein is the major component and the carbohydrate is the minor component, it is termed glycoprotein. © On the other hand, when the carbohydrate is the major component, and the protein is the minor component, it is termed proteoglycan.28 Medical Biochemistry: Preparatory Manual for Undergraduates Similarly, ina rate is the mi carbohydrate than 4%. A heteropolysaccharide containin, acid is known as mucopolysaccha: Glycoprotein is a protein’ contair (2-10 sugar residues) chains. Th glycolipid, the lipid is the major component, while the c inor component, Therefore, glycolipids group and itis said that the carbohydrate rbohy- are lipids containing a content in them is less ig repeating units of amino sugar and uronic ride/glycosaminoglycan. ning carbohydrate groups made up of short carbohydrate content is less than 4%. Proteoglycan is a heteropolysa ccharide joined to a polypeptide. Polysac- charides are the major component. The total carbohydrste content is over 90%. QI5. What are muc rides. Ans. polysaccharides? OR Write a note on mucopolysaccha- { Nucopolysaccharides are also called glycosaminoglycans (GAGs) (Fig. 3.14). * GAGs are complex, negatively charged heteropolysaccharides, which are normally found in association with small amounts of protein. mn CH,-0-|| xylose | [Gatecose anes) - [ Acidic _ Arie n sugar ~ sugar Serine Linker Fig. 3.14 Structure of glycosaminoglycans. Repeating units * These GAGs absorb lar; ge amount of water and which forms a basis of produce a geblike matrix, body's ground substances. The viscous, lubricati Broperties of mucous secretion are due to GAGs. Hence, they are called mu- copolysaccharides. * They help to maintain water and salt balance of the body. GAGs contribute to the ‘slippery’ property of mucous secretion, synovial fluid and vitreous hu- ron ora eve: The large negative charges cause the GAG chuine ne extend in solution and repel * The GAGs are capable of absorbin, l each other. and losing water very easily, GAGs act as effici : Fine us E *° The uronic acid is either glucuro) ronic nic acid or its C-5 epimer, Exception is keratan sulphate, Liduronic acid. instead of acidic in which galactose is present i sugar. * The amino sugar is either p- amino group is usually acetyl or C6. ose glucosamine or p-galactosamine, in which the lated. The amino sugar may be sulphated on C-4Chapter 3 — Chemistry of Carbohydrate © The combination of the sulphate and carboxylate groups gives the GAGs a very high density of negative charge. Some GAGs of biochemical importance are discussed below: (a) Hyaluronic acid * It isa viscous substance and a major component of the ground substance of connective tissue. * Itis also found in skin, synovial fluid, vitreous humour of eye, umbilical cord and also in bacteria. * Hyaluronic acid is composed of repeating units of p-glucuronic acid and N-acetylglucosamine (Fig. 3.15). On hydrolysis, it gives D-glucosamine, p-glucuronic acid and acetic acid. * It is the only GAG that is unsulphated and not attached to a protein. * Hyaluronic acid is hydrolysed by the enzyme hyaluronidase, which is present in sperm and testicular tissues. * Hyaluronidase hydrolyses the outer coat of the ovum, thus helping fertilisation. * Due to the action of this enzyme, viscosity of hyaluronic acid is reduced. Hence, it spreads in the tissues rapidly. © Hyaluronidase is therefore called ‘spreading agent’. It serves as a lubricant and shock absorber. CH,OH coo- H H A H OH H HO, H a H NHCOCH| H OH B-Glucuronic acid N-Acetylglucosamine tL —nh Fig. 3.15 Hyaluronic acid. (b) Chondroitin sulphate © Chondroitin sulphate (Fig. 3.16) is found in cartilage, bone, heart valves, tendons and cornea. om CH.OH coo" S2y H H HH H. NWor H He H H H NHCOCH, Hd Gal N-Acetyigalactosamine L___ B-Glucuronie acid sulphate, In Fig. 3.16 Chondroitin 4-sulphate.30 the C4 or the C6 position. Sho regulating a number of cellular activities. (©) Dermatan sulphate * Itis found in heart valves, blood vessels and skin. * Ghondroitin sulphates are the most abundant GAGs. * It forms proteoglycan aggregates, and in cartilage, 3 and holds the fibres tightly. Chondroitin sulphate is also involved in Medical Biochemistry: Preparatory Manual for Undergraduates © The structure of chondroitin is similar to that of hyaluronic acid. " * The difference is that the N-acetylglucosamine unit of the latter 1 replaced by an N-acetylgalactosamine, with the sulphate group on either it binds collagen * Dermatan sulphate contains l-iduronic acid and N-acetylgalactosamine. Small amounts of glucuronic acid are also found. Sulphate groups are present in C-4 positions. * Itis antithrombotic, but shows minimal anticoagulant activity. * Dermatan sulphate plays an important role in cardiovascular disease, wound repair, infection and fibrosis. (d) Heparin * Heparin occurs in combination with proteins as a proteoglycan. * It is an anticoagulant present mainly in liver and also in lungs, spleen, kidney and intestinal mucosa. * Heparin is composed of p-glucosamine-N-sulphate and sulphate ester of glucuronic acid linked by « (1-94) linkages (Fig. 3.17). Sulphate is ‘ found on C-3 and C-6 of glucosamine and C2 of uronic acid. The pro- residues. teoglycan of heparin is unique, consisting mainly of serine and glycine Fig. 3.17 Heparin. * Itis the only GAG with an a-linkage.. thrombin into thrombin. (e) Heparan sulphate groups. * It is involved in receptor signalling. (f) Keratan sulphate * It is found in cornea and tendons. * Heparin is also the only intracellular GAG. Asummary of GAGs is depicted in Table 3.4. CHSOs coo- A HOH ‘i ol NT 4H Z7Lo+N\ on A H H NHSOx Ht SOs" Sulphated glucosamine ‘Sulphated glucuronic acid * Heparin prevents blood clotting by inhibiting the conversion of pro- * This GAG is extracellular and forms a part of cell surfaces. * Similar to heparin, more acetylated groups are present than sulphate ¢ Ithelps in modulation of cellular activities with the environment. * Itis the only glycosaminoglycan that does not contain any uronic acid.Chapter 3 — Chemistry of Carbohydrates Chemical ‘Sulphation/ unique Tissue S. no. Glycosaminoglycan composition features distribution 1. Hyaluronic acid B-o-glucuronic Only GAG which Connective acid + is unsulphated tissue, skin, N-acetyiglucosamine and not attached. vitreous humour, B (153) to protein umbilical cord 2 Chondroitin sulphate -o-glucuronic acid + Sulphated in po- Cartilage, bones, N-acetyigalactosamine sitions 4 and 6 heart valve, 8 (13) Most abundant tendons, cornea GAG Heparin Glucuronic acid + Sulphated at Anticoagulant, D-glucosamine €-3orC-6of lungs, skin, mast a(i>4) glucosamine cells, intracellular and C-2 of GAG tronic acid 4. Heparan sulphate Glucuronic acid + Sulphated at_—_Lungs, blood N-acetyl- C-6 and vessel walls, o-glucosamine acetylated extracellular GAG (14) G&S Dermatan sulphate iduronic acid + Sulphated at Skin, tendons, N-acstylgalactosamine C-4 of galactos- heart valve, (133) amine and C-2 blood vessel of iduronic acid walls Iduronic acid present instead Of glucuronic acid 6 Keratan sulphate Galactose + Sulphated at Cartilage, cornea N-acetylglucosamine C-6 of both B (134) galactose and amino sugars Heterogeneous © The repeating units are galactose and N-acetylglucosamine with B-linkages (Fig. 3.18). It is the most abundant heterogeneous GAG. It can contain additional monosaccharides such as L-fucose and mannose. * It plays an important role in corneal transparency. © Itis involved in cellular recognition of protein ligands, cell motility and embryo implantation. CH,0©® 0H,0@) Q HO Q H H H H H OH H NHCOCH3 —— BrGalactose N-Acetyiglucosamine 1 Fig.3.18 Keratan sulphate.32 Medical Biochemistry: Preparatory Manual for Undergraduates (g) Blood group substances © Four monosaccharides are found in all types of blood group substances: galactose, fucose, acetylgalactosamine and acetylglucosamine. Nonreduc ing end groups of acetylglucosamine, galactose and fucose are associated with blood specificity of A, B and H, respectively. © The specific oligosaccharide chains are bound to proteins, lipids or membrane surfaces. Q\16. Write a note on mucopolysaccharidoses. Ans. © Mucopolysaccharidoses are hereditary disorders that are characterised by accu- mulation of GAGs in various tissues, e.g. Hunter syndrome, Hurler syndrome * Proteoglycans may act as autoantigens in various types of arthritis. © Chondroitin sulphate in cartilage is diminished with age, whereas keratan sulphate and hyaluronic acid are increased with age. © These changes may contribute to the development of osteoarthritis. Many dietary supplements used to treat osteoarthritis contain chondroitin sulphate. * Changes in GAGs also occur in skin on aging * Dermatan sulphate may play an important role in the development of athero- sclerotic plaque. © Hyaluronic acid may play an important role in facilitating tumour cells to migrate through the extracellular matrix. * Tumour cells induce the fibroblasts to synthesise increased amounts of hyaluronic acid. Table 3.5 gives the details. Products Other Deficient accumulated Clinical Syndromes names enzymes in urine manifestations Hurler-Scheie. MPS-1H a--iduronidase Keratan Comeal clouding, MPS-1S; sulphate mental retarda- autosomal Heparan tion, dwarf, | recessive sulphate deposition in coronary artery leads to ischaemia and early death Hunter MPS-Il; \duronate sulphatase Dermatan No corneal X-linked sulphate clouding, physical recessive Heparan deformity, mental sulphate retardation ~ mild to severe Sanfilippo A-D MPS-lIi, (a) Heparan, Heparan Severe nervous ‘autosomal sulphamidase sulphate system disorders, Tecessive _(b) N-acety| mental retardation glucosaminidase. (©) N-acetyl transferase (6) N-acetyl glucosamineProducts: | Other Deficient accumulated Clinical | Syndromes names enzymes in urine manifestations | Morquio. MPS-IV; (@) Galactose- Keratan Severe skeletal | AandB autosomal 6-Sulphatase ‘sulphate deformity spondy- recessive (b) B-Glucuronidase loepiphyseal dysplasia, no mental retardation | Maroteaux- MPS-VI; N-acetyl Dermatan Severe skeletal | Lamy autosomal —_(f-galactosamine ‘sulphate deformity, corneal recessive _sulphatase) no mental sly B-glucuronidase Dermatan. Hepatospleno- sulphate megaly, physical recessive Heparan deformity, mental sulphate retardation Q)7. Write a short note on glycoproteins. Ans. * Glycoproteins are proteins that have oligosaccharide chains covalently attached to their polypeptide backbones. © The carbohydrate content ranges from 1% to over 85%. * Glycoproteins have short oligosaccharide chains (1-20 sugars in length), which are highly branched, and they generally do not have a repeating sequence unlike proteoglycans, which have long, linear unbranched oligosaccharides with disaccharide repetitions. * Most of the plasma membrane proteins, blood proteins, receptors or hormones and blood group substances are glycoproteins. They serve as recognition sig- nals to facilitate cell—cell interactions, as antigens, as parts of the extracellular matrix and as mucins. © The principal sugars found in glycoproteins are galactose, glucose, mannose, fucose, N-acetylneuraminic acid (NANA), N-acetylgalactosamine (GalNAc) and N-acetylglucosamine (GlcNAc). The sugars may be attached to protein in either N-glycosidic or O-glycosidic linkage. The oligosaccharide chains are attached to the proteins via N- or O-glycosidic linkages. A glycoprotein is made up of only one type of these linkages or they have both in the same molecule. © In Nsinked glycosylation, the sugar is attached through a nitrogen (NH) within the asparagine or arginine side group on the protein. In O-linked glycosylation, the sugar is attached through oxygen on a hydroxyl group (OH) to a serine, threonine or tyrosine residue. © Ndinked oligosaccharides can be complex, high-mannose containing oligosac~ charides or hybrid types. © Niinked glycoproteins include such biologically important molecules as cell-surface receptors, cell-adhesion molecules, immunoglobulins and other serum proteins, and tumour antigens. * Oulinked oligosaccharides are found in the extracellular matrix or as part of membranes.34 Medical Biochemistry: Preparatory Manual for Undergraduates Q\18. Write a note on glycoprotein-related diseases. Ans. * Ininclusion-cell (I-cell) disease, lysosomal membrane proteins are not properly targeted to the lysosome because of the absence of a mannose 6-phosphate recognition signal owing to a genetically determined deficiency of N-acetyl glucosamine phosphotransferase. ; * In such people, lysosomes are deficient in certain hydrolases, do not function properly, accumulate partly digested cellular material and manifesting as inclusion bodies. * Deficiencies of specific glycoprotein lysosomal hydrolases such as mannosidases, cefucosidase, o-neuraminidase and aspartylglucosaminidase are genetically determined, resulting in abnormal degradation of glycoprotein. © The accumulation of glycoproteins in tissues can lead to various diseases. The four prominent diseases are mannosidosis, fucosidosis, sialidosis and aspartyl- glycosaminuria. * Major characteristics of these diseases are coarse facial features and mental re- tardation, vacuolisation of many cells and presence of abnormal degradation products in urine. Q.19. What are O-linked glycoproteins? Ans. * Olinked glycoproteins occur in mucous secretions (mucins). They are also found in some membranes and circulating glycoproteins. © The most common sugar attached is N-acetylgalactosamine. k * It is attached directly to serine (Ser) or threonine (Thr). It is this serine resi- due that is most likely to get phosphorylated as part of the signal transduction process. —2,6 NeuAc "==" _, GalNAc —____ Ser (Thr) © It presents a typical structure of an oligosaccharide chain present in O-linked glycoprotein. The synthesis of the O-linked glycoproteins uses nucleotide sug- ars such as uridine monophosphate (UDP) and cytidine monophosphate (CMP). * The enzymes catalysing the synthesis are glycoprotein glycosyltransferases. A glycosyl-galactose disaccharide may be linked to the hydroxyl group of hydroxylysine residues in collagen. * Only the Olinked glycoproteins remain in the mature collagen. The less gly- cosylated collagens tend to form fibrous structures, which occur in tendon, and the heavily glycosylated collagen is found in meshwork structures such as basement membranes. * In mucins, the O41 inked glycoproteins are usually short branched struc- tures containing N-acetylneuraminic acid (NANA) (sialic acids), galactose and N-acetylgalactosamine. N-acetylglucosamine and fucose may also be present. * The O-linked glycoproteins are negatively charged because of the presence of the sialic acid residues, and when they occur in clusters and in close proximity of each other, they repel each other. * Asa result, the protein assumes an extended state, yielding a highly viscous solution. Mucins form a protective barrier on the epithelial cell surface, lubricating and facilitating the transport process.Chapter 3 — Chemistry of Carbohydrates | Q.20. What are N-linked glycoproteins? Ans. * All the linked oligosaccharide chains found on proteins have branched structures having 2 common core structure of three mannose units and two N-acetylglucosamine residues (Fig. 3.19). * Beyond this core region, the N-linked glycoproteins are divided into three major classes. Complex © High mannose © Hybrid © Glycoproteins of the complex type are common in animals and generally have terminal trisaccharide sequences composed of sialic acid-galactose~GlcNAc attached to the core mannose structure. Fucose may be found in the core or in place of sialic acid. The oligosaccharide branches of the complex glycoproteins are often referred to as antennae, so that bi, tri, tetra- and penta-antennary structures may be found. © High-mannose oligosaccharides are found to a limited extent in animal glyco- proteins. They are more common in glycoproteins of lower eukaryotes and in viral envelop glycoproteins. © High-mannose oligosaccharides typically have two to six additional mannose residues linked to the pentasaccharide core. © Hybrid oligosaccharide molecules contain features of both the other two classes. © Biosynthesis of N-linked glycoproteins involves an oligosaccharide pyrophos- phoryl dolichol. Dolichols are polyprenols containing 16-20 isoprene units. Primary processes of degradation occur in the lysosomes, and the rest can occur in the endoplasmic reticulum. Man gy, @ S y Man B14) Gic Nac BUI>4) Gic NAc Asn 3) Man “0~* Fig. 8.19 Pentasaccharide core common to N-linked glycoproteins. POINTS TO PONDER sf © Carbohydrates can be synthesised via gluconeogenesis. Low-carbohydrate diets usually result in metabolic imbalances. Increasing the carbohydrate in the diet prevents the loss of nitrogen because of the ‘protein- sparing’ effect of carbohydrate. * Glucose is at the centre of carbohydrate metabolism and is the major dietary sugar. Diseases such as rheumatoid arthritis and osteoarthritis involve damage to molecules of the matrix.