Differential segmental flexibility and reach dictate the antigen binding mode of chimeric IgD and IgM: implications for the function of the B cell receptor

J Immunol. 2004 Mar 1;172(5):2925-34. doi: 10.4049/jimmunol.172.5.2925.

Abstract

Mature, naive B cells coexpress IgD and IgM with identical binding sites. In this study, the binding properties of such IgM and IgD are compared to determine how size and shape may influence their ability to bind Ag and thus function as receptors. To dissect their intrinsic binding properties, recombinant IgM and IgD were produced in soluble form as monomers of the basic H(2)L(2) Ab architecture, each with two Ag binding sites. Since these sites are connected with a hinge region in IgD and structural Ig domains in IgM, the two molecules differ significantly in this region. The results show that IgD exhibited the larger angle and longer distance between its binding sites, as well as having the greater flexibility. Relative functional affinity was assessed on two antigenic surfaces with high or low epitope density, respectively. At high epitope density, IgM had a higher functional affinity for the Ag compared with IgD. The order was reversed at low epitope density due to a decrease in the functional affinity of IgM. Studies of binding kinetics showed similar association rates for both molecules. The dissociation rate, however, was slower for IgM at high epitope density and for IgD at low epitope density. Taken together, the results show that IgM and IgD with identical Ag binding regions have different Ag binding properties.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibody Specificity / genetics
  • Binding Sites, Antibody* / genetics
  • Cell Line, Tumor
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / metabolism
  • Genetic Vectors
  • Immunoglobulin D / biosynthesis
  • Immunoglobulin D / genetics*
  • Immunoglobulin D / metabolism*
  • Immunoglobulin D / ultrastructure
  • Immunoglobulin M / biosynthesis
  • Immunoglobulin M / genetics
  • Immunoglobulin M / metabolism*
  • Immunoglobulin M / ultrastructure
  • Kinetics
  • Mice
  • Microscopy, Immunoelectron
  • Molecular Sequence Data
  • Nitrohydroxyiodophenylacetate / immunology*
  • Nitrohydroxyiodophenylacetate / metabolism
  • Receptors, Antigen, B-Cell / physiology*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / metabolism*
  • Recombinant Fusion Proteins / ultrastructure
  • Thiocyanates / chemistry
  • Transfection

Substances

  • Epitopes
  • Immunoglobulin D
  • Immunoglobulin M
  • Receptors, Antigen, B-Cell
  • Recombinant Fusion Proteins
  • Thiocyanates
  • Nitrohydroxyiodophenylacetate
  • thiocyanate