title: Characterization of C3-binding proteins on mouse neutrophils and platelets.
abstract: In the mouse, MCR1 and MCR2 on B lymphocytes are encoded by alternatively spliced Cr2 gene transcripts. Immune adherence receptors that bind C3 are present on mouse platelets and unstimulated neutrophils, yet they are not MCR1 or MCR2. To examine C3b- and C3d-binding proteins on mouse platelets and neutrophils, we performed C3b and C3d affinity chromatography as well as immunoprecipitation studies using previously described Ab to MCR1/MCR2 (mAb clones 8C12, 7G6, and 7E9 and polyclonal Ab BRN-1). Mouse neutrophils contained a 190-kDa membrane protein that specifically bound to C3b-Sepharose. Preabsorption of neutrophil proteins with anti-MCR1/MCR2 Ab did not affect the recovery of the 190-kDa C3b-binding protein by subsequent C3b affinity chromatography. Thus, this protein is immunologically distinct from the previously described MCR1 and MCR2 proteins. By virtue of its size and C3b-binding capacity, the 190-kDa protein was named C3bR-190. C3bR-190 was also apparent on platelets, but in reduced amounts. BRN-1 anti-MCR1/MCR2 Ab immunoprecipitated proteins of 125 and 150 kDa from surface-radiolabeled mouse platelets, which specifically bound to C3d-Sepharose. However, these proteins were not identified by mAb to MCR2, thus distinguishing them from previously described MCR2. These proteins were named C3dR-125 and C3dR-150. Therefore, we have identified a 190-kDa C3b-binding protein on mouse neutrophils and 125- and 150-kDa C3d-binding proteins on mouse platelets. These appear to be distinct from the heretofore identified mouse B lymphocyte MCR1 and MCR2. The identity of these C3b- and C3d-binding proteins on mouse neutrophils and platelets awaits further study.
authors.affiliation: Department of Medicine, The University of Chicago, IL 60637, USA.
authors.forename: R J
authors.initials: RJ
authors.lastname: Quigg
authors.forename: J J
authors.initials: JJ
authors.lastname: Alexander
authors.forename: C F
authors.initials: CF
authors.lastname: Lo
authors.forename: A
authors.initials: A
authors.lastname: Lim
authors.forename: C
authors.initials: C
authors.lastname: He
authors.forename: V M
authors.initials: VM
authors.lastname: Holers
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001792:Blood Platelets / Q000737:chemistry* / Q000378:metabolism; D002846:Chromatography, Affinity; D003176:Complement C3 / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D009504:Neutrophils / Q000737:chemistry* / Q000378:metabolism; D009928:Organ Specificity; D011233:Precipitin Tests; D017463:Receptors, Complement 3b / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D017464:Receptors, Complement 3d / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D003176:Complement C3; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d