Expression of immunoglobulin and scavenger receptor superfamily domains as chimeric proteins with domains 3 and 4 of CD4 for ligand analysis.
One approach to the analysis of leucocyte cell surface proteins is to express their domains with part of another protein as a carrier. We report the use of two immunoglobulin superfamily (IgSF) domains from rat CD4 (CD4d3 + 4) in producing domains from various superfamilies as chimeric proteins in Chinese hamster ovary cell lines. Four types of construct were successfully expressed containing: (i) the two IgSF domains of CD48; (ii) the IgSF domain of mb-1 which is part of the B cell antigen recognition complex; (iii) a T cell receptor V domain; and (iv) the N-terminal domain of CD5 which belongs to the scavenger receptor superfamily. This CD5 chimeric protein was antigenic for a panel of CD5 mAbs showing that mAbs with functional effects reacted with the N-terminal domain of CD5. The CD48 chimeric protein has been used both as multivalent complexes produced by cross-linking with mAbs recognizing CD4 and in a monomeric form to analyse the kinetics of the interaction between CD48 and CD2 [van der Merwe et al. (1993) EMBO J., 12, 4945-4954].
Brown
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MH
MRC Cellular Immunology Unit, Sir William Dunn School of Pathology, Oxford, UK.
Barclay
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AN
D000818:Animals; D000911:Antibodies, Monoclonal; D000917:Antibody Formation; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015704:CD4 Antigens / Q000096:biosynthesis / Q000235:genetics; D000071178:CD48 Antigen; D018956:CD5 Antigens; D051925:CD79 Antigens; D016466:CHO Cells; D006224:Cricetinae; D005822:Genetic Vectors; D006801:Humans; D007136:Immunoglobulins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D008024:Ligands; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D051381:Rats; D011947:Receptors, Antigen, B-Cell / Q000096:biosynthesis / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D051122:Scavenger Receptors, Class B
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015704:CD4 Antigens; D000071178:CD48 Antigen; C000604065:CD48 protein, human; D018956:CD5 Antigens; D051925:CD79 Antigens; C497881:CD79A protein, human; C000604066:Cd48 protein, mouse; C000604067:Cd48 protein, rat; D007136:Immunoglobulins; D008024:Ligands; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011947:Receptors, Antigen, B-Cell; D011948:Receptors, Antigen, T-Cell; D011971:Receptors, Immunologic; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D011993:Recombinant Fusion Proteins; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B