Activation of complement by an IgG molecule without a genetic hinge

Nature. 1993 Jun 17;363(6430):628-30. doi: 10.1038/363628a0.

Abstract

The hinge region links the two Fab arms to the Fc portion of the IgG molecule. It mediates flexibility to the molecule and serves as a connecting structure between the two heavy chains. In addition it provides space between the Fab and Fc parts. All three properties have been proposed to be important for the ability of IgG to initiate complement activation leading to complement-mediated cell lysis (CML). Here we report the construction of a hinge-deleted mouse-human chimaeric IgG3 molecule with specificity for the hapten NIP (3-iodo-4-hydroxy-5-nitrophenacetyl), HM-1. HM-1 lacks the genetic hinge, but has an introduced cysteine between Ala 231 (EU numbering) and Pro 232 in the lower hinge encoded by the CH2 exon. The introduced cysteine forms a disulphide bond between the two heavy chains of the molecule. In CML, HM-1 shows a greater activity than IgG3 wild type. This is the first time an IgG molecule without a genetic hinge has been found to be active in CML. We conclude that the hinge functioning as a spacer is not a prerequisite for complement activation. Rather, its major role seems to be to connect the heavy chains to each other in the amino-terminal part of CH2. Because HM-1 is expected to have low Fab-Fc flexibility, this molecular feature is probably of no importance for complement activation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cells, Cultured
  • Cloning, Molecular
  • Complement Activation / immunology*
  • Cysteine / chemistry
  • Cysteine / genetics
  • Cytotoxicity, Immunologic
  • DNA, Single-Stranded
  • Disulfides
  • Humans
  • Immunoglobulin Constant Regions / genetics
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology*
  • Immunoglobulin Heavy Chains / genetics
  • Mice
  • Molecular Sequence Data
  • Mutagenesis
  • Nitrohydroxyiodophenylacetate / immunology
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Restriction Mapping
  • Sheep
  • Structure-Activity Relationship

Substances

  • DNA, Single-Stranded
  • Disulfides
  • Immunoglobulin Constant Regions
  • Immunoglobulin G
  • Immunoglobulin Heavy Chains
  • Recombinant Fusion Proteins
  • Nitrohydroxyiodophenylacetate
  • Cysteine