The human complement system: assembly of the classical pathway C3 convertase

Biochem J. 1980 Jul 1;189(1):173-81. doi: 10.1042/bj1890173.

Abstract

The assembly of the classical pathway C3 convertase in the fluid phase has been studied. The enzyme is assembled from C2 and C4 on cleavage of these proteins by C1s. Once assembled, the enzyme activity decays rapidly. Kinetic evidence has been obtained that this decay is even more rapid than previously suggested (kdecay is 2.0 min-1 at 37 degrees C). As a result, optimal C3 convertase activity is only observed with high C1s levels, which result in rapid rates of cleavage of C2 and increased rates of formation of the C3 convertase. Using high concentrations of C1s at lower temperatures (22 degrees C) in the presence of excess substrate we have demonstrated kinetically that the enzyme comprises an equimolar complex of C4b and cleaved C2. We have obtained direct evidence from gel-filtration experiments for the role of C2a as the catalytic subunit of the enzyme. C2b appears to mediate the interaction between C4 (or C4b) and C2 at pH 8.5 and at low ionic strength where the interactions can easily be detected. It may therefore be important in the assembly of the enzyme, though it is not involved in the catalytic activity. The decay of the C3 convertase reflects the release of C2a from the C4b x (C2b) x C2a complex, and the stabilizing effect of iodine on the C3 convertase is therefore apparently one of stabilizing the C4b-C2z interaction, which is otherwise weak. C1s is not a part of the C3 convertase enzyme.

MeSH terms

  • Chemical Phenomena
  • Chemistry
  • Chromatography, Gel
  • Complement Activating Enzymes* / antagonists & inhibitors
  • Complement Activation*
  • Complement C2
  • Complement C3-C5 Convertases* / antagonists & inhibitors
  • Complement C4
  • Complement Pathway, Classical*
  • Humans
  • Isoflurophate / pharmacology
  • Kinetics

Substances

  • Complement C2
  • Complement C4
  • Isoflurophate
  • Complement Activating Enzymes
  • Complement C3-C5 Convertases