Glycosylation and stability of mature HIV envelope glycoprotein conformation under various conditions.
The role of the glycans of the mature human immunodeficiency virus (HIV) envelope (gp160) in its stability in various conditions was studied. gp160 conformation was monitored through its subsequent ability to bind [125I]CD4. Treatment of glycosylated (CHO+) gp160 with (i) sodium dodecyl sulfate (SDS) concentrations above 0.01% impaired subsequent CD4 binding while 0.3% SDS abolished it; (ii) beta-mercaptoethanol (MSH) concentrations above 0.01% impaired CD4 binding while 0.03% MSH abolished it; (iii) 2 M guanidine-HCl had no effect; (iv) temperatures between 50 degrees C and 80 degrees C altered CD4 binding while, above 80 degrees C, the binding was abolished; (v) CD4 binding was decreased by 50% by 2 freeze-thaw cycles but was not further affected by subsequent (up to 15) cycles; (vi) gp160 incubation in serum or cell lysate had no effect on CD4 binding. Glycanase treated (CHO-) gp160 binding activity was only 3-fold lower than that of CHO+ gp160. Only 2 M guanidine-HCl and heating at 70 degrees C differentially affected the binding of CHO+ and CHO- gp160, the effects being larger for CHO- gp160. CHO- gp160 binding was impaired after incubation in either serum or cell lysate. Thus, glycans stabilize gp160 conformation in some environments. However, CHO- gp160 appears to be resistant to denaturation as compared to other glycoproteins reported in the literature.
Papandreou
M J
MJ
IFR Jean Roche, URA 1455, Faculté de Médecine Nord, Marseille, France.
Idziorek
T
T
Miquelis
R
R
Fenouillet
E
E
D000818:Animals; D000911:Antibodies, Monoclonal; D015704:CD4 Antigens / Q000187:drug effects / Q000378:metabolism; D004355:Drug Stability; D015686:Gene Products, env / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D006031:Glycosylation; D006678:HIV / Q000276:immunology / Q000502:physiology*; D019302:HIV Envelope Protein gp160; D006801:Humans; D007700:Kinetics; D051379:Mice; D011485:Protein Binding; D011487:Protein Conformation; D011498:Protein Precursors / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D015686:Gene Products, env; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D012967:Sodium Dodecyl Sulfate