N-Acetyl-beta-D-glucosaminyl-binding properties of the envelope glycoprotein of human immunodeficiency virus type 1.
The effect of carbohydrate structures on the adsorption of HIV-1 or of recombinant envelope glycoprotein gp 160 (rgp 160) to cells of the CEM line was investigated with an indirect immunofluorescence assay using gp 120-specific mouse monoclonal antibodies (mAbs) directed to envelope gp 120. The beta-D-galactosyl, alpha-D-mannosyl, beta-D-glucosyl, N-acetyl-beta-D-glucosaminyl, sialosyl, and L-fucosyl derivatives tested had no effect on this binding. However, preincubation of HIV-1 (or rgp 160) with the neoglycoprotein, beta-D-GlcNAc47-BSA, specifically inhibited the labeling, by some of the mAb used, of HIV-1 (or rgp 160) bound at the cell membrane. This inhibition occurred only with mAbs that were specific for the immunodominant "neutralizing" third variable region (V3) of gp 120. Competition for the binding to rgp 160 between beta-D-GlcNAc47-BSA and mAb was further demonstrated by use of affinity matrices substituted with one of the relevant mAb (110-4), or with beta-D-GlcNAc47-BSA. Besides beta-D-GlcNAc47-BSA-Sepharose, rgp 160 also bound with low affinity, but high specificity, to two other N-acetyl-beta-D-glucosaminyl affinity matrices, beta-D-GlcNAc-divinylsulfone-agarose and asialoagalactothyroglobulin-agarose. Conversely, beta-D-[125I]GlcNAc47-BSA bound specifically to gp 160-Sepharose. These results indicated that rgp 160 behaves as a N-acetyl-beta-D-glucosaminyl-binding protein for GlcNAc residues presented at high density on a carrier, the carbohydrate-binding site of which is close to, or located on the V3 region of gp 120.
Laboratoire de Biologie Cellulaire, Faculté de Médecine Paris-Nord, Bobigny, France.
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Gattegno
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Sadeghi
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Saffar
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Bladier
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Clerget-Raslain
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JC
Gluckman
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Bahraoui
D000117:Acetylglucosamine / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D050260:Carbohydrate Metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D015686:Gene Products, env / Q000378:metabolism*; D015483:HIV Antibodies / Q000378:metabolism; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D015728:Receptors, HIV / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015686:Gene Products, env; D015483:HIV Antibodies; D019302:HIV Envelope Protein gp160; D037102:Lectins; D011498:Protein Precursors; D015728:Receptors, HIV; D012710:Serum Albumin, Bovine; D000117:Acetylglucosamine