3 D Structure of Immunoglobulins

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Presentation transcript:

3 D Structure of Immunoglobulins Hugh B. Fackrell Filename:Kuby05b7.ppt

Performance Objectives Assigned Reading Content Outline Performance Objectives Key terms Key Concepts Short Answer Questions

ASSIGNED READING Chapter pp

Antibody structure Special antibodies to study structure Myeloma proteins Bence Jones Proteins Monoclonal Antibodies 3D structure of antibodies Binding site Variability in structure

Angels & Devils

EM of IgA

Antibody flexibility

Antibody: Space filling model

Mulptiple Myeloma Disease Symptoms Detection Plasmocytoma Cancer of antibody producing cell Symptoms recurrent infections, anaemia, fractures, pain, bone lesions Detection Electrophoresis, X-rays, blood smear

IgG Crystal

Myeloma Induction

Bence Jones Protein Homogenous light chains In urine of myeloma patients Easily purified Insoluble at 60 -70 oC Soluble at 80 oC

Monoclonal Antibodies Add information here

Antibody model

Problems with flexibility Difficult to determine entire structure Hinge flexibility Fragment Study Fragment Assemble composite

Hinge Prolines

IgG: Enzyme hydrolysis

Bence Jones Light chains Simplest fragments to isolate Easily crystallized Mimics Fab

Bence Jones Monomer

Bence Jones Dimer

Immunoglobulin fold Signature structure common to all immunoglobulins 2 beta sheets held by disulfide bond Variations in detailed structure set specificity

Ig Fold: Variable domains

Ig fold: Constant domains

Antigen Binding Site

Structure of IgG

Ig Fold: hypervariable regions

Vitamin K bound to IgG

Antigen binding to Antibody

Antibody Strucutre: Internal motion

Antibody Structure: Internal movement

Antigen in Antibody

Hypervariable Regions Discovery KW plots Light chains Heavy chains CDR vs HVR

Kabat & Wu Plots Why Results demonstrate details of variability of protein sequence of similar proteins Results Variability = 1.0 >> no variation Variability = 400 >> 20 different AA Number different amino acids Variability = Frequency of most common AA

HV: light chains

KW plot heavy chains

HV:IgG

KW Heavy chains CDR

KW Light chains CDR

Ig Superfamily

Isotypes Allotypes Idiotypes

Allosera Production

Isotypes, Allotype Idiotype

The End

Performance Objectives Key terms, concepts short answers

SHORT ANSWER QUESTIONS