Purification of a lamprey complement protein homologous to the third component of the mammalian complement system.
A lamprey protein homologous to the third component of mammalian complement was isolated from lamprey plasma and was tentatively designated lamprey C3. Lamprey C3 is a major protein in lamprey serum with electrophoretic mobility of beta-globulin and with m.w. of 190,000. It consists of three polypeptide chains (84,000-alpha, 74,000-beta, and 32,000-gamma chains) linked by disulfide bonds. The protein retains a unique internal thiolester bond on the alpha-chain that is cleaved on methylamine treatment or on limited proteolysis with trypsin. Lamprey C3 by itself could not bind to zymosan or rabbit red cells, but it could covalently bind to these substances when activated by other factors present in lamprey serum. The binding of lamprey C3 to activating surfaces is mediated by covalent bonds and is accompanied by limited proteolysis of the alpha-chain. A fragment with an m.w. of 35,000 containing the internal thiolester site was isolated from methylamine-treated lamprey C3 bound to activated thiol-Sepharose by extensive tryptic digestion followed by dithiothreitol treatment. Lamprey C3 functions as the essential factor in phagocytosis of rabbit red cells by lamprey phagocytes. However, it is not involved in naturally occurring hemolytic activity in lamprey serum.
D000818:Animals; D001665:Binding Sites; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003176:Complement C3 / Q000302:isolation & purification* / Q000378:metabolism / Q000502:physiology; D003602:Cytotoxicity, Immunologic; D005399:Fishes / Q000276:immunology*; D007798:Lampreys / Q000276:immunology*; D010446:Peptide Fragments / Q000032:analysis; D010587:Phagocytosis; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000302:isolation & purification; D015054:Zymosan / Q000378:metabolism
D003176:Complement C3; D010446:Peptide Fragments; D013438:Sulfhydryl Compounds; D015054:Zymosan