Coreceptor function of mutant human CD4 molecules without affinity to gp120 of human immunodeficiency virus.
Despite extensive mutational studies on the human CD4 molecule and its affinity to human immunodeficiency virus (HIV) envelope glycoprotein gp120, coreceptor functions of such mutant molecules have only been examined by indirect measurement of their affinity to class II major histocompatibility complex (MHC) molecules. In this report, coreceptor functions of mutant human CD4 molecules, which have no or reduced affinity to an HIV envelope protein, gp120, were assessed in a murine T cell receptor/class II MHC recognition system. The substitution of human C" beta strand with the murine homologous segment resulted in the loss of the coreceptor function as well as in the complete loss of gp120 binding capacity, corroborating the consensus that Phe-43 in C" beta strand plays crucial roles in both situations. However, simultaneous replacement of the C'-C" loop along with the C" beta strand by homologous murine segments rescued the coreceptor function, whereas gp120 binding capacity remained negative. Further analysis indicated that insertion of lysine between Gly-41 and Ser-42 can partially compensate for the coreceptor function lost by the Phe-43 --> Val mutation. Although the coreceptor function of these mutant CD4 molecules in a human T cell recognition system is yet to be determined, these observations necessitate a re-evaluation of the role played by Phe-43 in coreceptor function. Examination of the sensitivities of the mutant CD4 molecules expressed on HeLa cells to infection by a T cell-tropic HIV-1 strain indicated that only those mutants that had completely lost gp120 binding capacity were resistant to the infection. All mutants having whole C" substitution, irrespective of additional substitutions or their coreceptor functions, were resistant to the infection.
Tachibana
M
M
Department of Immunology, Mitsubishi Kasei Institute of Life Sciences, Machida, Tokyo 194, Japan.
Siddiqi
M A
MA
Ikegami
Y
Y
Eshima
K
K
Shirota-Someya
Y
Y
Tahara-Hanaoka
S
S
Koito
A
A
Iizuka
M
M
Shinohara
N
N
D000595:Amino Acid Sequence; D000818:Animals; D015704:CD4 Antigens / Q000235:genetics* / Q000276:immunology; D005434:Flow Cytometry; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology* / Q000472:pathogenicity; D006367:HeLa Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology; D014162:Transfection
D015704:CD4 Antigens; D015699:HIV Envelope Protein gp120; D011948:Receptors, Antigen, T-Cell