Isolation and characterization of a novel plasma protein which binds to activated C4 of the classical complement pathway

We report here the isolation and partial characterization of a previously unrecognized protease-sensitive plasma protein identified during the development of a novel protocol for the purification of the second component of human complement (C2). This new protein is physicochemically similar to C2. I...

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Veröffentlicht in:The Journal of biological chemistry 1989-02, Vol.264 (4), p.2283-2291
Hauptverfasser: Hammer, C H, Jacobs, R M, Frank, M M
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Jacobs, R M
Frank, M M
description We report here the isolation and partial characterization of a previously unrecognized protease-sensitive plasma protein identified during the development of a novel protocol for the purification of the second component of human complement (C2). This new protein is physicochemically similar to C2. It coprecipitates with C2 on polyethylene glycol fractionation and specifically binds, like C2, to Sepharose-bound iC4/C4b. Binding occurs both in the presence and absence of C2. The purified protein has a chain structure similar to C2 as determined by sodium dodecyl sulfate-gel electrophoresis in the presence or absence of reducing agent and has a molecular mass of 120 kDa, only somewhat greater than C2 at 95 kDa. Both proteins radioiodinate under similar conditions to the same specific activities with each of two different methods that yield 10-fold disparate results. Quantitative Mancini analysis identifies 300 µg/ml of the 120-kDa protein in plasma and serum. The protein is present at normal concentrations in serum from individuals genetically deficient in C2, has no C2 functional activity, and is not cleaved as is C2 when serum complement is activated. Potent monospecific polyclonal anti-serum to each do not cross-immunoprecipitate using standard gel techniques. However, these anti-sera identify epitopes in common by Western blotting. The data presented indicate that the 120-kDa protein is a distinct plasma component and suggest that the protein is not an “immature” form of C2. Initial experiments to delineate a functional role for the 120-kDa protein have demonstrated a consistent inhibition of C1 site generation on EAC4b which is dose-dependent and reversible. Thus, this protein appears to be a new complement regulatory factor.
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Potent monospecific polyclonal anti-serum to each do not cross-immunoprecipitate using standard gel techniques. However, these anti-sera identify epitopes in common by Western blotting. The data presented indicate that the 120-kDa protein is a distinct plasma component and suggest that the protein is not an “immature” form of C2. Initial experiments to delineate a functional role for the 120-kDa protein have demonstrated a consistent inhibition of C1 site generation on EAC4b which is dose-dependent and reversible. 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Potent monospecific polyclonal anti-serum to each do not cross-immunoprecipitate using standard gel techniques. However, these anti-sera identify epitopes in common by Western blotting. The data presented indicate that the 120-kDa protein is a distinct plasma component and suggest that the protein is not an “immature” form of C2. Initial experiments to delineate a functional role for the 120-kDa protein have demonstrated a consistent inhibition of C1 site generation on EAC4b which is dose-dependent and reversible. 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Psychology</topic><topic>Humans</topic><topic>Immune Sera</topic><topic>Immunodiffusion</topic><topic>Immunoelectrophoresis</topic><topic>Molecular Weight</topic><topic>Protein Binding</topic><topic>Proteins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hammer, C H</creatorcontrib><creatorcontrib>Jacobs, R M</creatorcontrib><creatorcontrib>Frank, M M</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hammer, C H</au><au>Jacobs, R M</au><au>Frank, M M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation and characterization of a novel plasma protein which binds to activated C4 of the classical complement pathway</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1989-02-05</date><risdate>1989</risdate><volume>264</volume><issue>4</issue><spage>2283</spage><epage>2291</epage><pages>2283-2291</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>We report here the isolation and partial characterization of a previously unrecognized protease-sensitive plasma protein identified during the development of a novel protocol for the purification of the second component of human complement (C2). 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subjects Analytical, structural and metabolic biochemistry
Binding and carrier proteins
Biological and medical sciences
Blood Proteins - isolation & purification
Blood Proteins - metabolism
Chromatography, Affinity
Chromatography, Ion Exchange
Complement Activation
Complement C4 - metabolism
Complement Pathway, Classical
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
Humans
Immune Sera
Immunodiffusion
Immunoelectrophoresis
Molecular Weight
Protein Binding
Proteins
title Isolation and characterization of a novel plasma protein which binds to activated C4 of the classical complement pathway
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