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. Author manuscript; available in PMC: 2009 Nov 1.

Published in final edited form as: Structure. 2008 Nov 12;16(11):1689–1701. doi: 10.1016/j.str.2008.09.005

a. The binding of [³H]-NBD-556 to equivalent amounts of the w.t. or S375W HIV-1_YU2 gp120 glycoprotein or bovine serum albumin (BSA) is indicated. The means from a single experiment, with the standard errors derived from triplicate samples, are shown. The experiment was performed three times with comparable results. b. The amount of radiolabeled w.t. or S375W mutant HIV-1_YU2 gp120 bound to Cf2Th-CCR5 cells is shown, after incubation without (0) or with CD4-Ig (1 μg/ml), NBD-556 (100 μM) or BMS-806 (10 μM). Numbers indicate molecular weight (in kD). c. Recombinant viruses bearing the w.t. or indicated mutant HIV-1_YU2 envelope glycoproteins were incubated with Cf2Th-CCR5 cells in the presence of increasing concentrations of NBD-556. Luciferase activity in the Cf2Th-CCR5 cells was measured, and the relative virus infectivity, compared with that observed in the absence of added NBD-556, is shown. The mean values and standard errors (from triplicate samples) are shown from a typical experiment. d. The location of HIV-1_YU2 gp120 residues in which changes affect NBD-556 binding and/or entry enhancement is shown. The modeled NBD-556, with bonds colored according to the identity of the atoms (see Figure 3 legend), is shown penetrating into the Phe 43 cavity of gp120. The gp120 core is depicted as a C_α ribbon and transparent molecular surface. The HIV-1_YU2 gp120 residues that, when changed, affect NBD-556 binding and/or enhancement of HIV-1 entry into Cf2Th-CCR5 cells are labeled and highlighted in arbitrary colors, with side chain bonds shown.

a. The binding of [³H]-NBD-556 to equivalent amounts of the w.t. or S375W HIV-1_YU2 gp120 glycoprotein or bovine serum albumin (BSA) is indicated. The means from a single experiment, with the standard errors derived from triplicate samples, are shown. The experiment was performed three times with comparable results. b. The amount of radiolabeled w.t. or S375W mutant HIV-1_YU2 gp120 bound to Cf2Th-CCR5 cells is shown, after incubation without (0) or with CD4-Ig (1 μg/ml), NBD-556 (100 μM) or BMS-806 (10 μM). Numbers indicate molecular weight (in kD). c. Recombinant viruses bearing the w.t. or indicated mutant HIV-1_YU2 envelope glycoproteins were incubated with Cf2Th-CCR5 cells in the presence of increasing concentrations of NBD-556. Luciferase activity in the Cf2Th-CCR5 cells was measured, and the relative virus infectivity, compared with that observed in the absence of added NBD-556, is shown. The mean values and standard errors (from triplicate samples) are shown from a typical experiment. d. The location of HIV-1_YU2 gp120 residues in which changes affect NBD-556 binding and/or entry enhancement is shown. The modeled NBD-556, with bonds colored according to the identity of the atoms (see Figure 3 legend), is shown penetrating into the Phe 43 cavity of gp120. The gp120 core is depicted as a C_α ribbon and transparent molecular surface. The HIV-1_YU2 gp120 residues that, when changed, affect NBD-556 binding and/or enhancement of HIV-1 entry into Cf2Th-CCR5 cells are labeled and highlighted in arbitrary colors, with side chain bonds shown.