The Immunoglobulins Structure and Function

Visit the link below to download the full version of this book:

https://medidownload.com/product/the-immunoglobulins-structure-and-function/

Click Download Now

 Contents

Preface xi

I PA RT
Structural Aspects

1 G e n e r a l C h a r a c t e r i s t i c s of I m m u n o g l o b u l i n M o l e c u l e s
I. Immunoglobulin Classes 4
A. Immunoglobulin G 5
B. Immunoglobulin M 6
C. Immunoglobulin A 7
D. Immunoglobulin D 9
E. Immunoglobulin E 9
II. Heterogeneity of Immunoglobulins 10
III. Detection and Isolation of Immunoglobulins 11
A. Detection of Immunoglobulins 11
B. Isolation of Immunoglobulins 12
IV. Proteolytic and Chemical Fragments of Immunoglobulins 13
A. Fragmentation by Proteinases 13
B. Chemical Fragmentation 16
V. Immunoglobulin Peptide Chains 17
A. General Features 17
B. Variable Regions 19
C. Constant Regions 21
VI. Additional Peptide Chains 28
A. J Chain 28
B. Secretory Component 29
vi Contents

C. Surrogate Chains on the B-Cell Precursors 31

D. Accessory Peptide Chains of the B-Cell Antigen
Receptor Complex 32
VII. Immunoglobulin Fold (Domain) 36
A. Structural Characteristics 36
B. Domains of Members of Immunoglobulin Superfamily 38
C. Contacts between Domains 40
VIII. Fab Portion 42
IX. Fc Portion 43
X. Immunoglobulin Molecules 43
A. Dissociation and Reassociation of Immunoglobulin
Peptide Chains 43
B. Intact Immunoglobulin Molecules 46
XI. Carbohydrate Components of Immunoglobulins 52
A. Linkage of Glycans to Immunoglobulin Peptide Chains 53
B. Chemical Structure of Immunoglobulin Glycans 54
C. Distribution of Glycans on Immunoglobulin Molecules 55
D. Three-Dimensional Studies of Immunoglobulin
Glycans 57
E. Functions of Immunoglobulin Carbohydrates 59
F. Variations of Galactosylation 60
References 62

2 Animal and Human Immunoglobulins

I. Low Vertebrates 76
A. Fishes 76
B. Amphibians 79
C. Reptiles 8O
II. Birds 81
A. Chickens 81
B. Ducks 82
III. Mammals 84
A. Laboratory Animals 84
B. Farm Animals 91
C. Pets 94
IV. Human Immunoglobulins 95
A. General Considerations 95
B. Human Allotypes 96
C. Human Immunoglobulin Genes 98
V. Origin of Antibody Diversity in Mice and Humans 100
A. V(D)J Recombination 102
B. Somatic Hypermutation 103
C. Class Switching 104
Contents vii

VI. Evolutionary Aspects 105

A. Immunoglobulin-Type Domains in Prokaryotes
and Invertebrates 105
B. Evolution of Immunoglobulin Domains 110
References 112

3 E n g i n e e r i n g A n t i b o d y Molecules
I.
Problems of Serum Immunotherapy 123
II.
Chimeric Antibody Molecules and Humanization 125
III.Minimal Antibody Fragment (Fv) 127
IV.Production of Human Monoclonal Antibodies
by Phage-Display and Transgene Technologies 130
A. Phage-Display Technology 130
B. Transgenic Animals 133
V. Engineering of Immunoglobulins with Novel Properties 134
A. Fusion Proteins 134
B. Immunotoxins 136
VI. Polymerization of IgG Molecules and Their Fragments 137
VII. Bispecific Antibodies 139
References 142

PART I I
Functional Aspects

4 Antigen-Combining Site
I. General Characteristics 151
A. Canonical Conformations of Hypervariable Loops 152
B. Correlation of Binding Site Surface with Type of Antigen 161
C. Bonds between Antigen and Antibody 164
D. Residues Responsible for Contacts with Antigens 165
E. Water Molecules Participate in Antigen-Antibody Interactions 168
II. Conformational Changes Linked with Antigen Binding 169
A. Induced Fit 170
B. Global Changes 171
III. Complex of VH Domain of Camelid Heavy Chain Antibodies
with Antigen 172
IV. Interaction of an Autoantibody (Rheumatoid Factor)
with Fc 7 173
viii Contents

V. Structural Aspects of Antibody Specificity 174

A. Complexes of a Single Antibody with Several
Related Antigens 174
B. Complexes of a Single Epitope with Different Antibodies 176
C. Effect of Mutations on the Antigen-Combining Site 177
D. Structure of Idiotype--Anti-Idiotype Complexes 179
VI. Modeling Antibody-Combining Sites 182
A. Knowledge-Based Methods 184
B. ab initio Procedures 185
C. Application of Modeling 186
VII. Structural Aspects of Catalytic Antibody Activity 189
A. Catalytic Antibodies Induced by Immunization 189
B. Autoantibodies with Catalytic Activity 193
References 195

5 Antigen-Recognizing Molecules O t h e r T h a n A n t i b o d i e s
I. T-Cell Antigen Receptor 205
II. Proteins of Major Histocompatibility Complex 207
III. Complexes of T-Cell Receptor and Peptide-MHC 211
IV. CD1 Molecules 212
V. Natural Killer Cell Inhibitory Receptors 213
VI. Comparison of Antigen Binding by Various
Antigen-Recognizing Molecules 215
References 216

6 I n t e r a c t i o n s O u t s i d e the A n t i g e n - C o m b i n i n g Site
I. Fc Receptor-Binding Sites 221
A. Binding Sites for Fc Receptors Involved in Effector Responses 223
B. Binding Sites for Fc Receptors Involved in Transcytosis 227
II. Complement-Binding Sites 230
A. C lq Binding 231
B. C3b and C4b Binding 234
C. C3a, C4a, and C5a Anaphylatoxin Binding 234
III. Proteins Reacting with the Fc Portion 235
A. Clusterin 235
B. Human Plasma Glycoprotein 60 235
C. Fibronectin 235
D. Fc-Binding Peptides 236
E. Seminal Proteins 236
E G-Actin 237
G. Placental Alkaline Phosphatase 237
H. Herpes Simplex Virus Proteins 237
Contents ix

IV. Proteins Reacting with the Fab Portion 237

A. Prolactin 237
B. Protein Fv 238
C. T-Cell Protein CD4 238
D. HIV Protein gp120 239
V. Lectins 239
VI. Molecular Chaperones 241
VII. Bacterial Immunoglobulin-Binding Proteins 243
A. Staphylococcal Protein A 243
B. Streptococcal Protein G 245
C. Protein H 247
D. Protein L 247
References 247

7 Segmental Movements of Immunoglobulin Molecules

I. General Aspects 255
II. Functional Aspects of Segmental Flexibility 259
References 261
Index 263
This Page Intentionally Left Blank
 Preface

Modern immunology evolved from revolutionary discoveries that occurred a

hundred years ago, during the last decade of the 19th century. In 1890, Emil
Behring and Shibasaburo Kitasato observed that sera obtained from rabbits pre-
viously injected with bacterial toxins could neutralize those toxins. Further-
more, they found that the injection of these sera into other animals conferred a
long-term resistance to the toxins, and thus could be used as an effective thera-
peutic agent. These findings provided solid evidence that blood-born sub-
stances can mediate immune responses to toxins. Soon afterward, Paul Ehrlich
showed that sera obtained from rabbits injected with plant toxins also had
toxin-neutralization activity. The neutralizing activity of antitoxin sera was cor-
related by Ehrlich with a distinct group of macromolecules. Ehrlich hypothe-
sized that cells have surface receptors with an affinity for a particular antigen.
Upon detachment of the surface receptors from cells, the receptors could neu-
tralize toxins. The high specificity of antitoxins was attributed by Ehrlich to
structural complementarity between chemical groupings on the antitoxins and
toxins. Based on the chemical concepts, he developed a quantitative method for
the detection of antitoxins and toxins as chemical reactants. All these pioneer-
ing studies and novel concepts led to the birth of a new discipline, termed
"immunochemistry" by Svante Arrhenius.
During the following decades, these concepts were corroborated and ex-
panded. Karl Landsteiner, Michael Heidelberger, and other immunochemists
showed that not only toxins from plants or bacteria can elicit specific im-
munological responses but also polysaccharides and a seemingly unlimited
number of chemical substances (haptens). In the 1930s, the antigen-specific
macromolecules in serum were described as a particular class of serum proteins,
which were named antibodies. Elvin Kabat during his work in the laboratories
of Arne Tiselius and Theodor Svedberg showed that the antibody activity of

xi
xii P~eface

rabbit antiserum was due to 7S gammaglobulins. This was achieved by subject-

ing the antiserum to adsorption on the immunizing antigen, which depleted a
significant portion of the 7S gammaglobulins from the immune serum. These
classic experiments of Kabat initiated studies of antibodies at the molecular level.
A turning point occurred in 1958-1959 with the studies of Rodney Porter
and Gerald Edelman. Porter developed a mild procedure for proteolysis of rab-
bit immunoglobulin G (IgG) using papain and succeeded in separating two
large proteolytic fragments. One, the Fab fragment, can interact with antigens
and contains one antigen-binding site. The other, a crystallizable Fc fragment,
cannot combine with antigen, but has other important biological activities.
Concurrently, Edelman separated the human IgG1 molecule into constituent
subunits, the large (heavy) and small (light) peptide chains, by reduction of
interchain disulfide linkages. In 1962, Porter suggested his famous structural
model, according to which each IgG molecule is composed of four polypep-
tide chains (two identical heavy and two identical light chains) joined by
disulfide bonds.
Due to the rapid development of molecular immunology in the 1960s and
1970s, the main features of immunoglobulin structure and biosynthesis were
elucidated, as were the genetic mechanisms responsible for the enormous vari-
ability of antibodies. This tremendous body of knowledge about antibodies was
presented in several books of that period (Kabat, 1968; Pressman and Gross-
berg, 1968; Nisonoff et al., 1975; Nezlin,1977).
During the 1980s and 1990s, the accumulation of knowledge about immuno-
globulins has continued at an accelerated rate. Precise structural data on how
the antibody-combining site interacts with antigens has come from x-ray crys-
tallographic studies and has advanced modeling of the antibody-combining
sites. Intense effort has been directed toward furthering our understanding
of the genetic mechanisms of antibody formation, which has facilitated the gen-
eration of new types of antibody molecules in vitro. In fact, some of these new
antibodies have already been effectively used in biotechnology and medicine.
Achievements in molecular immunology have and continue to be published
in a continous flow of original papers and reviews. This monograph provides
one source in which recent concepts regarding both immunoglobulin structure
and function have been summarized. Structural and functional properties of
major immunoglobulin classes of human and animal imunoglobulins are
treated at length. The localization and the structure of different binding sites of
immunoglobulin molecules, including first of all the antigen-binding site, are
discussed on the basis of latest x-ray crystallography studies. Emphasis has
been placed on recently solved problems and their practical applications. Re-
cent reviews are cited in each section where additional information and exten-
sive relevant citations can be obtained for topics not extensively covered herein.
Even with the tremendous advances in understanding the structure and func-
Preface xiii

tion of immunoglobulins, achieved during the last century, there are still
enough mysteries concerning the imunoglobulin molecules remaining to be
solved, which presents an exciting challenge to us all.

REFERENCES

Kabat E.A. (1968). Structural Concepts in Immunology and Immunochemistry. Holt, Rinchart and
Winston, New York.
Pressman D., and Grossberg A.L. (1968). The Structural Basis of Antibody Specificity. WA. Ben-
jamin, New York.
NisonoffA., HopperJ.E., and Spring S.B. (1975). The Antibody Molecule. Academic Press, New York.
Nezlin R. (1977). Structure and Biosynthesis of Antibodies. Plenum Press, New York.
This Page Intentionally Left Blank
 PART I

Structural Aspects
This Page Intentionally Left Blank
 CHAPTER 1
General Characteristics of
Immunoglobulin Molecules

Immunoglobulin molecules of all classes are heterodimers and composed from

four polypeptide chains--two identical large or heavy (H) chains of about
50-60 kDa and two small or light (L) chains of about 23 kDa, linked by inter-
chain disulfide bridges (Fig. 1). This prototype structure, first suggested by
Rodney Porter in 1962, is common for all monomeric immunoglobulin mole-
cules. Polymeric immunoglobulins of higher molecular weights are formed by
2-6 four-chain subunits, similar to the monomeric immunoglobulin molecules.
They possess one or two additional peptide chains, which are important for
formation and stabilization of immunoglobulin polymers. All immunoglobulin
molecules are glycoproteins and contain two or more carbohydrate chains, usu-
ally linked to heavy chains. Membrane forms of immunoglobulins serve as a
specific part of the B-cell antigen receptors. They have additional peptide
stretches on the COOH-terminal ends of their heavy chains, with which they
are embedded in cell membranes, as well as short portions of chains inside the
cell (intracellular segments of different sizes). Membrane immunoglobulins are
associated noncovalently with two accessory peptides forming the B-cell anti-
gen receptor complex in the B lymphocyte membrane.
 Immunoglobulin Molecules

N N

H
Z CDR

Fab L
C

Hinge
I
m
C2 |
t)

JH
Fc
C3 !1)i 1 o

C
FIGURE 1 The basic four-chain structure of immunoglobulin molecules. H and L, the heavy and
light peptide chains; N and C, NH 2- and COOH-terminal ends of the peptide chains; VL and VH,
variable regions of the light and heavy chains; C1, C2, and C3, homology regions (domains) of the
heavy chains; CDR, complementarity-determining region. The distribution of interchain and in-
trachain disulfide bonds are shown. Shadowed ovals point to localization of oligosaccharides in CH2
domains. (Nezlin, 1994. Reprinted with permission from Marcel Dekker, NY.)

I. I M M U N O G L O B U L I N CLASSES

In humans and rodents, there are five immunoglobulin classes or isotypes,

which differ in the primary structure, carbohydrate content, and antigenic
properties of their heavy chains (Table 1). By contrast, the light chain types are
the same for all immunoglobulin classes. Each immunoglobulin molecule con-
tains light chains of one of two types, either lambda (2) or kappa (x). The 2 and
light chains have different primary structures and antigenic properties. They
are usually free of carbohydrate components. The ratio of x/2 chains in human
and swine immunoglobulins is about 60:40, whereas in mouse, rat and rabbit
immunoglobulins it is about 95:5. Some other mammals such as dog, cat, and
farm animals (ox, sheep, and horse) have mainly 2 chains and chicken im-
munoglobulins contain only 2 chains.