title: The second component of human complement: use of glycosidases and glucosylation to distinguish the two forms.
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 degrees C resulted in a four- to fivefold increase in the hemolytic activity of both forms. The increase in activity was different than 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 degrees 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.
authors.affiliation: Department of Medicine, University of Miami School of Medicine, Florida.
authors.forename: D R
authors.initials: DR
authors.lastname: Schultz
authors.forename: M
authors.initials: M
authors.lastname: Loos
mesh_terms: D003175:Complement C2 / Q000302:isolation & purification / Q000378:metabolism*; D006026:Glycoside Hydrolases; D006031:Glycosylation; D006801:Humans; D007455:Iodine; D007700:Kinetics; D009439:Neuraminidase
chemical_list: D003175:Complement C2; D007455:Iodine; D006026:Glycoside Hydrolases; D009439:Neuraminidase