PMID: 6977086Dec 4, 1981Paper

Isolation of three separate anaphylatoxins from complement-activated human serum

Molecular and Cellular Biochemistry
T E HugliS Russell

Abstract

Recent methodologies used in preparing anaphylatoxins from complement-activated serum are described. Activation of the alternative pathway generates C3a and C5a; however, activation of the classical pathway is required to generate the anaphylatoxin from C4. This article describes an activation scheme that simultaneously generates all three of the anaphylatoxins (e.g., C3a, C4a and C5a) in human serum and outlines a procedure for isolating each as homogeneous products. Purification of intact anaphylatoxins directly from complement-activated serum takes place only if an exopeptidase in serum, known as carboxypeptidase N (SCPN), is properly inhibited. A new series of mercapto derivatives of arginine analogs are introduced as potent and effective inhibitors of SCPN. These inhibitors permit normal complement activation but prevent degradation of the released activation fragments C3a, C4a or C5a. The SCPN inhibitor previously used was 6-aminohexanoic acid (EACA), but it required a 1 M concentration for effective inhibition, the substituted mercapto-guanido compounds prove to be effective in the mM range.

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Citations

Sep 1, 1993·Protein Science : a Publication of the Protein Society·D F Carney, T E Hugli
Aug 1, 1994·Protein Science : a Publication of the Protein Society·L CuiT E Hugli
Jun 1, 1988·Inflammation·H R LamcheD E Hammerschmidt
Jun 1, 1983·Molecular Immunology·T E HugliB W Erickson
Jul 1, 1988·Molecular Immunology·L CuiT E Hugli
Dec 1, 1984·Immunopharmacology·R HueyT E Hugli
Feb 1, 1987·Immunopharmacology·V G Nielsen, R O Webster
Dec 1, 1983·Journal of Neuroimmunology·N SchupfJ Cox
Jan 1, 1987·Journal of Neuroimmunology·N Schupf, C A Williams
Mar 1, 1989·Brain, Behavior, and Immunity·N SchupfL Kerper
Jun 9, 2004·Developmental and Comparative Immunology·Yoko KatoTomoki Yano
Feb 26, 1998·Immunopharmacology·J F Regal
Feb 26, 1998·Immunopharmacology·R S AmesH M Sarau
Jul 1, 1985·Journal of Neuroimmunology·C A WilliamsT E Hugli
Dec 20, 1996·Cancer Letters·G Hetland, T E Hugli
Jul 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·D G NettesheimE R Zuiderweg
Sep 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·S C BischoffC A Dahinden
Aug 1, 1989·The Journal of Experimental Medicine·Y KurimotoC A Dahinden
Dec 14, 1999·Protein Science : a Publication of the Protein Society·J SunT E Hugli
Jul 1, 1983·The Journal of Clinical Investigation·C A DahindenT E Hugli
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Nov 23, 2005·Microbiology and Immunology·Masayoshi AbeHidechika Okada
May 1, 1996·The Journal of Laboratory and Clinical Medicine·R J JohnsonD E Van Epps
May 1, 1984·The Journal of Surgical Research·J K HornM R Flick
Dec 1, 1985·British Journal of Haematology·M A Lett-Brown, H S Juneja

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