The second component of human complement: Use of glycosidases and glucosylation to distinguish the two forms

Duane R. Schultz, Michael Loos

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

The two forms of human plasma C2 that were described in the preceding report (1) were investigated for their functional and biochemical differences. Incubation with the neuraminidase (NAN'dase) of Clostridium perfringens at 37°C resulted in a four- to fivefold increase in the hemolytic activity of both forms. The increase in activity was different from the increase caused by treatment with iodine. The mechanism of increased activity of NAN'dase-treated C2 was the generation of increased molecules of activated C3 (C3b), resulting in more molecules of C5 binding to (C4b,2a,3b)(n). Removal of N-acetyl-neuraminate from C2 did not alter its binding to a cationic exchanger. Nonenzymatic glucosylation was used to distinguish the two forms of C2. Incubation of highly pure C2 with 14C-D-glucose resulted in the gradual accumulation of radioactivity in acid-precipitable material. The two forms of C2 were glucosylated in vitro for seven days with 14C-D-glucose in phosphate-buffered saline at 25°C. Form 2 bound twice as much 14C-D-glucose as form 1. Glucosylated form 2, but not form 1, lost some of its affinity to bind to a cationic exchanger. Since the interaction between glucose and protein occurs at free amino groups, we conclude that form 2 of C2 has approximately twice as many free amino groups as form 1. This explains the reason for the existence of two forms of C2 in plasma independent of the allelic variant.

Original languageEnglish (US)
Pages (from-to)394-409
Number of pages16
JournalImmunobiology
Volume176
Issue number4-5
DOIs
StatePublished - Jan 1 1988

Keywords

  • DGVB
  • EDTA
  • ELISA
  • GlcNAc'dase
  • N-acetylglucosaminidase
  • NAN'dase
  • enzyme-linked immunosorbent assay
  • ethylenediaminetetraacetate
  • gal'dase
  • galactosidase
  • glucose gelatin-Veronal with 0.15 mM Ca 2 and 0.5 mM Mg, relative NaCl concentration 75 mM, pH 7.5
  • neuraminidase

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Hematology

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