Characterization of physiologic breakdown products of the complement fragment Ba.
To better characterize the activation products of factor B which are generated under physiologic conditions Ba was purified directly from human EDTA-plasma by immunoaffinity chromatography using anti-Ba Sepharose. SDS-PAGE analysis revealed the existence of degradation products of the Ba fragment which were truncated at the carboxyterminus. A monoclonal antibody (mAb D22/3) was produced by immunizing mice with a synthetic peptide which corresponds to the Ba carboxyterminus (Glu215-Arg234). This mAb was found to react with an epitope (Ba neo-epitope), which is newly formed after the generation of Ba from its precursor protein factor B. This neoantigenic determinant is absent both in factor B and the desArg/Lys Ba derivatives. The conversion of Ba by carboxypeptidases in human serum was monitored using an assay which is based on mAb D22/3, revealing a half-life of Ba in serum of 150 min. Furthermore, this assay allowed to quantitate plasma levels of intact and degraded Ba in healthy probands and in patients with chronic renal failure. The processing of the Ba carboxyterminus may be of functional relevance as the biological activity of the Ba fragment which had been shown to suppress human B lymphocyte functions in vitro resides in its carboxyterminal amino acid sequence.
Oppermann
M
M
Department of Immunology, University of Göttingen, Germany.
Götze
O
O
D000595:Amino Acid Sequence; D043424:Carboxypeptidase B; D002268:Carboxypeptidases / Q000494:pharmacology; D011415:Complement Factor B / Q000276:immunology / Q000378:metabolism*; D003170:Complement Pathway, Alternative; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D051437:Renal Insufficiency / Q000378:metabolism; D014511:Uremia / Q000378:metabolism
D010446:Peptide Fragments; D004492:Edetic Acid; D002268:Carboxypeptidases; D043424:Carboxypeptidase B; D011415:Complement Factor B