Identification of functional domains on gC1Q-R, a cell surface protein that binds to the globular ''heads'' of C1Q, using monoclonal antibodies and synthetic peptides
Identification of functional domains on gC1Q-R, a cell surface protein that binds to the globular ''heads'' of C1Q, using monoclonal antibodies and synthetic peptides
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DOI：
10.1089/hyb.1996.15.333
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发表时间：
1996-10-01
期刊：
影响因子：
--
通讯作者：
Peerschke, EIB
中科院分区：
文献类型：
--
作者：
Ghebrehiwet, B;Lu, PD;Peerschke, EIB
A membrane protein (33 kDa) that binds to the globular ''heads'' of Clq (gC1q-R) has been recently described, The full length cDNA encoding gC1q-R has been cloned, expressed in E. coli and using the purified recombinant protein (rgC1q-R) as an immunogen, a panel of IgG monoclonal antibodies (MAb) has been produced by fusion of spleen cells from hyperimmunized BALB/c mice with NSO mouse myeloma partners, From this fusion, 60 anti-gC1q-R hybridomas were selected and evaluated for their ability to (1) discriminate between the mature form (MF) of gC1q-R (residues 74-282) and a truncated form (TF) lacking residues 74-95, which contains a major Clq binding site, (2) recognize two functionally defined synthetic peptides derived from the NH2-(XN(18)) and COOH-(XC(15)) terminus of gC1q-R, and (3) bind to microtiter well fixed intact Raji cells, Several clones were identified: MAbs 46.23 and 60.11 (IgG(1 kappa), reacted strongly with ELISA plate-fixed intact Raji and K562 cells, MF, and the XN(18) peptide, but had poor or no reactivity with TF; MAbs 74.5.2 > 25.15 (IgG(1 kappa)) recognized both MF and TF and are directed against epitopes in the XC(15) peptide that contains a binding site for high-molecular-weight kininogen and Factor XII.
