Interaction of human C1q with IgG and IgM: revisited

Biochemistry. 2008 Dec 9;47(49):13093-102. doi: 10.1021/bi801131h.

Abstract

The first step of activation of the classical complement pathway involves the binding of the globular C1q domain (gC1q) to the antigen-bound IgG or IgM. To improve our understanding of the mechanism of interaction of gC1q with IgG and IgM, we compared the immunoglobulin binding properties of single-residue mutants of individual globular modules of A and C chains. We found that Lys(A200) and Lys(C170) are significant for binding with both immunoglobulins. In addition, two C1q-specific scFv antibodies known as potent inhibitors of C1q-IgG and -IgM interactions were used in the epitope mapping analysis. A set of important residues, which participate in the C1q epitopes for scFv, were identified: Lys(C170) for the scFv3(V) epitope and Arg(B108) and Arg(B109) for the scFv10(V) epitope. The ability of scFv3(V) and scFv10(V) to bind preformed C1q-IgG or C1q-IgM complexes differed: scFv3(V) retained its ability to bind C1q, while scFv10(V) lost it. Given the different locations of the epitopes and the varying abilities of both antibodies to bind C1q-IgG and C1q-IgM complexes, we found that residues from the apical surface of C1q [where the scFv3(V) epitope was located] were involved in the initial recognition of IgG and IgM, while Arg(B108) and Arg(B109) are able to interact during the initial recognition as well as during the final binding of immunoglobulins. The reported results provide the first experimental evidence supporting the notion that apical and equatorial surfaces of gC1q have consecutive involvement following the gC1q reorientation during the interaction with specific C1q ligands.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arginine / chemistry
  • Arginine / genetics
  • Arginine / metabolism
  • Binding Sites
  • Complement C1q / chemistry
  • Complement C1q / genetics
  • Complement C1q / metabolism*
  • Epitopes / genetics
  • Epitopes / immunology
  • Epitopes / metabolism
  • Humans
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / genetics
  • Immunoglobulin G / metabolism*
  • Immunoglobulin M / chemistry
  • Immunoglobulin M / genetics
  • Immunoglobulin M / metabolism*
  • Ligands
  • Lysine / chemistry
  • Lysine / genetics
  • Lysine / metabolism
  • Protein Binding
  • Protein Conformation

Substances

  • Epitopes
  • Immunoglobulin G
  • Immunoglobulin M
  • Ligands
  • Complement C1q
  • Arginine
  • Lysine