Dimerization of CD4-C kappa chimeric molecules leads to loss of CD4 epitopes.
Structural similarities between two members of the immunoglobulin superfamily were explored by making chimeric immunoglobulin/CD4 antigen molecules. A crossover in the middle of the originally proposed J kappa homology unit of the first domain of the CD4 molecule was used to construct a chimeric molecule having human and mouse CD4 antigen sequence through the first 108 amino acids and murine J kappa and C kappa sequence thereafter. This molecule was expressed in the presence and absence of an immunoglobulin heavy chain. The resulting proteins were assayed for the expression of CD4 epitopes that should be present based on epitope mapping data. Monomeric, homodimeric, and heavy chain/light chain tetrameric forms of the recombinant protein were secreted and were all detectable with anti-kappa reagents. CD4 antibodies precipitated only the form of the CD4-C kappa light chain protein which appears as a monomer by polyacrylamide gel electrophoresis. Neither the homodimer nor the heavy chain/light chain tetramer were detected with CD4 monoclonal antibodies. An engineered gene having this CD4 antigen first domain joined to the human IgG1 constant region, when coexpressed with a mouse lambda light chain, also failed to express detectable CD4 epitopes. The structural implications of the presence or absence of CD4 epitopes on these proteins is discussed.
Frey
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Becton Dickinson Monoclonal Center, San Jose, CA 95131.
Estess
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D000818:Animals; D001483:Base Sequence; D015704:CD4 Antigens / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D000939:Epitopes; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000737:chemistry* / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000737:chemistry* / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D014162:Transfection
D015704:CD4 Antigens; D000939:Epitopes; D007127:Immunoglobulin Constant Regions; D007145:Immunoglobulin kappa-Chains; D011993:Recombinant Fusion Proteins