CD4-binding of gp130 micelles isolated from SIVagmTYO-7.
The binding and binding inhibition of the SIVagmTYO-7 external glycoprotein gp130 in micellar form to the CD4 molecule on human Molt-4 clone 8 cells was investigated. The best binding of gp130 to Molt-4 clone 8 cells occurred at pH 5.5 to 6.5 at 37 degrees C after 4 h or at room temperature after 10 h. The dissociation constant of this reaction was 0.2-0.4 nM, with both soluble CD4 or CD4 on Molt-4 clone 8 cells. This value is close to 0.15 nM determined for the antihuman CD4 monoclonal antibody 30F16H5. After partial deglycosylation of gp130, a 90 kD product arose which still bound to CD4. Fully deglycosylated gp130 of 60 kD was still immunoprecipitable, but had lost the CD4 binding activity. Lens culinaris agglutinin was able to inhibit the gp130-CD4 interaction very efficiently, while the agglutinin of Phaseolus vulgaris was half as efficient and Canavalia ensiformis was inefficient. CD4 binding of gp130 micelles was also inhibited with several anti CD4 monoclonal antibodies directed against the OKT4a epitope as well as with soluble recombinant CD4.
German Primate Centre, Kellnerweg, FRG.
F
F
Polzien
J G
JG
Scharf
W
W
Lüke
G
G
Hunsmann
D000911:Antibodies, Monoclonal / Q000276:immunology; D001667:Binding, Competitive; D015704:CD4 Antigens / Q000276:immunology / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008823:Micelles; D015302:Simian Immunodeficiency Virus / Q000378:metabolism*; D013696:Temperature; D014759:Viral Envelope Proteins / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D008823:Micelles; D014759:Viral Envelope Proteins