CICLOS FUTILES

Los ciclos ftiles, que se podran producir entre las reacciones irreversibles situadas al mismo nivel de la glucolisis y la gluconeognesis, consumiran ATP. En general no se producen fisiolgicamente gracias a la regulacin conjunta de ambas vas: glucoltica y gluconeognica. Si los consideramos como ciclos de sustrato, sirven para amplificar las seales metablicas y para producir calor.

576

Chapter 15

Principles of Metabolic Regulation: Glucose and Glycogen

Glucose
hexokinase glucose 6-phosphatase

Glucose 6-phosphate
phosphohexose isomerase

Fructose 6-phosphate
phosphofructokinase-1 fructose 1,6-bisphosphatase

Fructose 1,6-bisphosphate Dihydroxyacetone phosphate

triose phosphate isomerase aldolase

Dihydroxyacetone phosphate
triose phosphate isomerase

Glycolysis

(2) Glyceraldehyde 3-phosphate

glyceraldehyde phosphate dehydrogenase

Gluconeogenesis

(2) 1,3-Bisphosphoglycerate
phosphoglycerate kinase

(2) 3-Phosphoglycerate
phosphoglycerate mutase

(2) 2-Phosphoglycerate
enolase

(2) Phosphoenolpyruvate
PEP carboxykinase pyruvate kinase

(2) Oxaloacetate
pyruvate carboxylase

(2) Pyruvate

FIGURE 1515 Glycolysis and gluconeogenesis. Opposing pathways of glycolysis (pink) and gluconeogenesis (blue) in rat liver.

(the bypass reactions) and glycolysis; seven steps are catalyzed by the same enzymes in the two pathways. Cofactors have been

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15.3

Coordinated Regulation of Glycolysis and Gluconeogenesis

Gluconeogenesis

582

Chapter 15

Principles of Metabolic Regulation: Glucose and Glycogen Fructose 6-phosphate ATP

ATP ADP
FBPase-1 activity (% of Vmax)

Pi

100

PFK-1

PFK-1 activity (% of Vmax)

AMP citrate

100

FBPase-1

O O B B O OP O O O OP O OOCH2 A A O O O H HO H CH2OH OH H
Fructose 2,6-bisphosphate

80 60

F26BP

When fructose 2,6-bisphosphate binds to its allo site on PFK-1, it increases that enzymes affinity f Glycolysis substrate, fructose 6-phosphate, and reduces its 40 40 ity for the allosteric inhibitors ATP and citrate. A FIGURE 1521 Regulation of fructose 1,6-bisphosphatase-1 (FBPase-1) physiological concentrations of its substrates ATP F26BP and phosphofructokinase-1 (PFK-1). The important 20 20 role of fructose F26BP fructose 6-phosphate and of its other positive and 2,6-bisphosphate in the regulation of this substrate cycle is detailed ative effectors (ATP, AMP, citrate), PFK-1 is vir in subsequent figures. 0 0 in the absence of fructose 2,6-bisphosp 0 0.05 0.1 0.2 0.4 0.7 1.0 2.0 4.0 0 50 inactive 100 Fructose 2,6-bisphosphate activates PFK-1 and s [Fructose 6-phosphate] (mM) [Fructose 1,6-bisphosphate] ( M) lates glycolysis in liver and, at the same time, inh (a) in the two pathways are regulated in a coordinated and (b) FBPase-1, thereby slowing gluconeogenesis. Gluconeogenesis reciprocal manner. In general, when sufficient concenAlthough structurally related to fructose trations of acetyl-CoA or citrate (the product of acetylbisphosphate, fructose 2,6-bisphosphate is not a CoA ATP condensation with oxaloacetate) are present, or Pi Fructose 6-phosphate termediate in gluconeogenesis or glycolysis; it is a when a high proportion of the cells adenylate is in the ulator whose cellular level reflects the level of gluc form of ATP, gluconeogenesis is favored. AMP promotes PFK-1 F26BP FBPase-1 in the blood, which rises when blood glucose falls glycogen degradation and glycolysis by activating glycocellular concentration of fructose 2,6-bisphospha gen phosphorylase (via activation of phosphorylase kiset by the relative rates of its formation and break Fructose 1,6-bisphosphate ADP nase) and stimulating the activity of PFK-1. H2O (Fig. 1523a). It is formed by phosphorylatio All the regulatory actions discussed here are trigfructose 6-phosphate, catalyzed by phosphofruc gered by changes inside the cell and are mediated by Glycolysis nase-2 (PFK-2), and is broken down by fructose (c) very rapid, instantly reversible, allosteric mechanisms. bisphosphatase (FBPase-2). (Note that thes Another set of regulatory processes is triggered from FIGURE 1522 Role of fructose 2,6-bisphosphate in regulation of 0.08 mM. Thus F26BP activates PFK-1 by increasing apparent affinzymes its are distinct from PFK-1 and FBPase-1, w outside the cell by the hormones insulin and glucagon, glycolysis and gluconeogenesis. Fructose 2,6-bisphosphate (F26BP) ity (Fig. 1518) for fructose 6-phosphate. (b) FBPase-1 activity is incatalyze the formation and breakdown, respective which signal too much or too little glucose in the blood, has opposite effects on the enzymatic activities of phosphofructokihibited by as little as 1 M F26BP and is strongly inhibited by 25 M. fructose 1,6-bisphosphate.) PFK-2 and FBPaserespectively, or by epinephrine, which signals the imnase-1 (PFK-1, a glycolytic enzyme) and fructose 1,6-bisphosphatase 1,6- activities of a single, bifunc In the absence of this inhibitor (blue curve)two the K 0.5 for fructose distinct enzymatic pending need for fuel for a fight-or-flight response. These (FBPase-1, a gluconeogenic enzyme). (a) PFK-1 activity in the absence bisphosphate is 5 M, but in the presence ofprotein. 25 M F26BP curve) of these two activities in the The(red balance hormonal signals bring about covalent modification of F26BP (blue curve) is half-maximal when the concentration of the K0.5 is 70M. Fructose 2,6-bisphosphate also determines makes FBPase-1 which the cellular level of fructose (phosphorylation or dephosphorylation) of target profructose 6-phosphate is 2 mM (that is, K0.5 2 mM). When 0.13 M more sensitive to inhibition by another allosteric regulator, AMP. (c) bisphosphate, is regulated by glucagon and insulin teins inside the cell; this takes place on a somewhat longer
60 F26BP is present (red curve), the K for fructose 6-phosphate is only Summary of regulation by F26BP.

ADP

80 Fructose 1,6-bisphosphate

F26BP H2O

Glycolysis

(c)

FIGURE 1522 Role of fructose 2,6-bisphosphate in regulation of

glycolysis and gluconeogenesis. Fructose 2,6-bisphosphate (F26BP) has opposite effects on the enzymatic activities of phosphofructokinase-1 (PFK-1, a glycolytic enzyme) and fructose 1,6-bisphosphatase (FBPase-1, a gluconeogenic enzyme). (a) PFK-1 activity in the absence of F26BP (blue curve) is half-maximal when the concentration of fructose 6-phosphate is 2 mM (that is, K0.5 2 mM). When 0.13 M F26BP is present (red curve), the K0.5 for fructose 6-phosphate is only

0.08 mM. Thus F26BP activates PFK-1 by increasing its apparent affinity (Fig. 1518) for fructose 6-phosphate. (b) FBPase-1 activity is inhibited by as little as 1 M F26BP and is strongly inhibited by 25 M. In the absence of this inhibitor (blue curve) the K0.5 for fructose 1,6bisphosphate is 5 M, but in the presence of 25 M F26BP (red curve) the K0.5 is 70M. Fructose 2,6-bisphosphate also makes FBPase-1 more sensitive to inhibition by another allosteric regulator, AMP. (c) Summary of regulation by F26BP.

[F26BP] Stimulates glycolysis, inhibits gluconeogenesis ATP

PFK-2

PFK-2 (active) FBPase-2 (inactive)

OH

Fructose 6-phosphate
FBPase-2

Pi
insulin

Pi
phosphocAMP-dependent protein protein kinase phosphatase

ATP
glucagon ( [cAMP])

ADP

Fructose 2,6-bisphosphate

H2O PFK-2 (inactive) FBPase-2 (active) O O P O O

ADP

(a)
[F26BP] Inhibits glycolysis, stimulates gluconeogenesis

(b)

FIGURE 1523 Regulation of fructose 2,6-bisphosphate level. (a) The

cellular concentration of the regulator fructose 2,6-bisphosphate (F26BP) is determined by the rates of its synthesis by phosphofructokinase-2 (PFK-2) and breakdown by fructose 2,6-bisphosphatase

(FBPase-2). (b) Both enzymes are part of the same polypeptide chain, and both are regulated, in a reciprocal fashion, by insulin and glucagon. Here and elsewhere, arrows are used to indicate increasing (h) and decreasing (g) levels of metabolites.

Metabolic Regulation: Glucose and Glycogen

glycogen synthase activity. as three Ser residues near its ylated by glycogen synthase synthase to the inactive (b) phosphorylation (priming) by ation of glycogen synthase b e pathway for this action in rotein phosphatase (PP1 in n muscle, epinephrine actiycogen-targeting protein GM ciation of PP1 from glycogen. ylation of glycogen synthase ation that is a good substrate sphorylation; the binding of ces a conformational change en phosphorylase b, thus re29).

Insulin

Phosphoserines near carboxyl terminus P

3ADP GSK3

3ATP

ADP CKII ATP

P P

Glycogen synthase b Inactive

HO HO HO Glycogen synthase a Active

PP1

3Pi

Insulin

Glucagon, epinephrine

Glucose 6-phosphate

Glucose

lated

a glucose 6-phosphate sensor. In muscle, a different

Ciclo de CORI
Consiste en un acoplamiento de dos rutas metablicas (glucolisis y gluconeognesis) en dos rganos distintos (msculo e hgado), que permite a las clulas musculares poder disponer de la energa necesaria en todo momento. El msculo obtiene ATP a partir de la degradacin de glucosa en la glucolisis. Cuando las condiciones del ejercicio son anaerbicas la glucosa se degrada a piruvato y ste se reduce a lactato. El lactato es exportado a la circulacin y es captado por el hgado. El hgado sintetiza glucosa de nuevo a partir de lactato por la ruta gluconeognica.

Ciclo de Cori

Ciclo Glucosa/Alanina
Glucosa Glucosa

2 ATP

Urea

6 ATP

(2) Piruvato
4 ATP

(2) Piruvato

(2) Lactato
aminocido

(2)-NH2

Enzima?

-Ceto cido

(2) Alanina

(2) Alanina

Sangre
Hgado Msculo

En general involucra AA transferasas y GPT moviendo nitrgeno desde el msculo al hgado. GPT: Glutamato piruvato transaminasa. Note la similitud con el ciclo de Cori!!

ations of the two aminotransferases by the SGPT GOT tests (S for serum)and of another encreatine kinase, by the SCK testcan pro-

ful in the monitoring of people exposed to these chemicals, because these enzyme activities are high in liver and can be detected in very small amounts.

Transports Ammonia from l Muscles to the Liver

Muscle protein Amino acids NH 4 Glucose
glycolysis

also plays a special role in transporting amino to the liver in a nontoxic form, via a pathway he glucose-alanine cycle (Fig. 189). In muscertain other tissues that degrade amino acids , amino groups are collected in the form of te by transamination (Fig. 182a). Glutamate converted to glutamine for transport to the liver, ribed above, or it can transfer its -amino group uvate, a readily available product of muscle is, by the action of alanine aminotransferase 9). The alanine so formed passes into the blood vels to the liver. In the cytosol of hepatocytes, aminotransferase transfers the amino group anine to -ketoglutarate, forming pyruvate and te. Glutamate can then enter mitochondria, he glutamate dehydrogenase reaction releases ig. 187), or can undergo transamination with etate to form aspartate, another nitrogen donor synthesis, as we shall see. e use of alanine to transport ammonia from muscles to the liver is another example of the c economy of living organisms. Vigorously conskeletal muscles operate anaerobically, producuvate and lactate from glycolysis as well as

Pyruvate

Glutamate

alanine aminotransferase

Alanine

-Ketoglutarate

Blood glucose

Blood alanine

Alanine

-Ketoglutarate
Glutamate

alanine aminotransferase

Glucose

gluconeogenesis

Pyruvate

NH 4
urea cycle

89 Glucose-alanine cycle. Alanine serves as a carrier of

and of the carbon skeleton of pyruvate from skeletal musr. The ammonia is excreted and the pyruvate is used to proose, which is returned to the muscle.

Liver

Urea

Integracin Metablica y Vas Unidireccionales

Temas
Perfiles Metablicos Anlisis del Metabolismo Unidireccionalidad Metabolismo en un Organismo Multicelular

Perfiles en el metabolismo de combustible

Tejido Cerebro Msculo esq (reposo) Msculo esq (ejercicio) Msculo Cardiaco Tejido adiposo Hgado Combustible almacenado Ninguno Glucgeno Combustible preferido Glucosa (cuerpos cetnicos) Acidos grasos Combustible exportados Ninguno Ninguno

Ninguno

Glucosa

Lactato, Alanina

Ninguno

Acidos grasos

Ninguno

Triacilgliceroles Glucgeno triacilgliceroles

Acidos grasos Aminocidos glucosa, cidos grasos

Acidos grasos, glicerol Acidos grasos, glucosa cuerpos cetnicos

Principales hormonas que controlan metabolismo de combustible

Hormona Insulina Acciones bioqumicas permeabilidad celular a glucosa (musc. y Tej. Adip.) Gluclisis Sntesis de glucgeno Sntesis de triacilgliceroles Gluconeognesis Liplisis Degradacin de protenas Sntesis de protenas, DNA y RNA Concentracin de cAMP (hgado y tej. adiposo) Glucogenlisis Sntesis de glucgeno Hidrlisis de triacilgliceroles Gluconeognesis Gluclisis Concentracin de cAMP en msculo Movilizacin de triacilgliceroles Glucogenlisis Sntesis de glucgeno Acciones fisiolgicas Seala el estado de ingesta Concentracin de glucosa en sangre Almacenamiento de combustible Crecimiento y diferenciacin celular

Glucagn

Liberacin de glucosa del hgado Concentracin de glucosa en sangre

Adrenalina

Liberacin de glucosa del hgado Uso de glucosa por el msculo Concentracin de glucosa en sangre

Anlisis del Metabolismo

Vas Catablica y anablica, ocurren simultneamente, deben actuar de manera regulada, ordenadamente, respuesta total
Solo uno pocos intermediarios conectan sistemas principales - azcar-Ps, alfa-ceto cidos, derivados CoA, y PEP ATP & NADPH relacionan catabolismo & anabolismo

Mecanismos frecuentes de regulacin metablica

Anabolismo y catabolismo deben coordinarse con precisin
Interacciones alostricas Modificacin covalente Niveles enzimticos Compartimentalizacin Especializacin metablica en los rganos

Interacciones alostricas
Los posibles puntos de control son enzimas que catalizan reacciones irreversibles
Fosfofrutoquinasa (gliclisis): Activadores AMP, ADP, fructosa 2,6 bifosfato. Inhibidores ATP y citrato

Acetil CoA carboxilasa (sntesis de cidos grasos): Activador citrato. Inhibidor palmitoil CoA

Modificacin Covalente
Algunas enzimas son controladas por modificacin covalente, adems de interaccin alostrica
Glucgeno fosforilasa: dos formas (fosforilasa a [activa] y fosforilasa b [relativamente inactiva]. Fosforilacin de Ser14, aumenta actividad de la enzima. Glucgeno sintasa: dos formas (sintasa I [activa] y sintasa D [menos activa]. La enzima disminuye su actividad cuando se fosforila.

Estequiometra Metablica
Tres tipos de estequiometra en sistemas biolgicos Reaccin estequiomtrica - el nmero de cada tipo de tomo en una reaccin Estequiometra de apareamiento obligada requiere el acoplamiento de transportadores de electrones Estequiometra acoplada el nmero de molculas de ATP en la vas involucra comsumirlo o producirlo -

Naturaleza de equivalente de ATP

Una perspectiva diferente G para hidrlisis de ATP dice que a concentraciones de equilibrio de ADP y Pi deberan ser muy superiores a ATP Sin embargo, una clula donde esto sea cierto, es porque est muerta Control cintico sobre las vas catablicas asegura que la razn [ATP]/[ADP][Pi] permanezca muy alto Esto permite que hidrlisis de ATP puede servir como la fuerza conductora para la mayora de los procesos bioqumicos

El ATP equivalente
Cul es el coeficiente de acoplamiento" para producir o consumir ATP? Coeficiente de acoplamiento son los moles de ATP producidos o consumidos por mol de substrato convertido (o producto formado) Oxidacin celular de glucosa tiene un coeficiente de acoplamiento de 30-38 (dependiendo del tipo de clula) Hexoquinasa tiene un coeficiente de acoplamiento de -1 Piruvato kinasa (en gliclisis) coeficiente de acoplamiento de +1

El significado de los 38 ATPs

La estequiometra del ATP" tiene un gran efecto sobre la Keq de una reaccin Considere la Keq para la oxidacin de glucosa Si se producen 38 ATP, el G celular es -967 kJ/mol y Keq = 10170, un nmero muy grande! Si G = 0, se pueden hacer 58 ATP, pero la reaccin estar en equilibrio con solo la mitad de la glucosa oxidada que nosotros tenemos De forma que el nmero de 38 es un compromiso!

Significado de grandes Keq

A mayor ATP generado, ms baja la constante de equilibrio, y ms alto el nivel de glucosa requerida Si [glucosa] esta bajo este valor, no ser efectivamente utilizada Grandes Keq significa que los niveles basales de glucosa sern muy bajos Grandes Keq tambin significan que la reaccin estar lejos del equilibrio y puede ser regulada

El valor ATP de NADH vs NADPH

El valor ATP para NADH es 2.5-3 El valor ATP para NADPH es mayor NADPH transporta electrones desde vas catablicas a procesos biosintticos [NADPH]>[NADP+] por ello NADPH/NADP+ es mejor sistema donante de e- que NADH/NAD De manera que NADPH promueve 3.5-4 ATP!

Vas unidireccionales
Un rol "secreto" de ATP en metabolismo Ambas direcciones de alguna va opuesta debe ser favorable, de forma que el efector alostrico puede controlar efectivamente la direccin El coeficiente de acoplamiento de ATP para alguna de esas secuencias ha evolucionado de manera que todo el equilibrio para la conversin es altamente favorable

Combustible para el cerebro

Cerebro tiene un alto metabolismo pero no tiene combustible de reserva Esto significa que el cerebro requiere un suplemento constante de glucosa En condiciones extremas, cerebro puede usar -hidroxibutirato (desde cidos grasos), convirtiendolo en acetil-CoA en TCA Esto permite que el cerebro use grasas como combustible!

Creatina Kinasa en Msculo

Msculos deben ser preparados para una rpida provisin de energa Creatina kinasa y fosfocreatina actan como sistema tampn, proveyendo ATP adicional para la contraccin Glicgeno provee energa adicional, liberando glucosa para la gliclisis Gliclisis baja rpidamente pH, causando fatiga muscular

Fosfocreatina

Creatinina

Degradacin de la protena msculo

Durante alta actividad, los amino cidos se degradan a piruvato, los cuales pueden ser transaminados a alanina Alanina circula al hgado, donde es convertido nuevamente a piruvato alimento para gluconeognesis Este es el ltimo recurso para organismos exhaustos o que presentan alta actividad

Piruvato

Gluatamato

Glutamato alanina aminotransferasa

-cetoglutarato Alanina

Integracin del Metabolismo

Urea Consumo de Protenas Acidos grasos

NH3

Acetil CoA

Protenas corporales

AA AA no escenciales Energa CO2 + H2O

Purinas Pirimidinas Porfirinas

Neurotransmisores Fosfolpidos enzimas

Productos intermediario Gliclisis y ciclo de Krebs

Glucosa + depsito glicgeno