Journal:

Article Title: Biochemical and Genetic Characterizations of a Novel Human Immunodeficiency Virus Type 1 Inhibitor That Blocks gp120-CD4 Interactions

doi: 10.1128/JVI.77.19.10528-10536.2003

Figure Lengend Snippet: Competitive inhibition of gp120-CD4 binding by BMS-378806 and binding affinity of BMS-378806 to gp120. (A) Saturation binding curve of BMS-378806. The levels of sCD4 bound to gp120 in the absence or presence of various concentrations of BMS-378806 and in escalating concentrations of sCD4 were measured by using the gp120-CD4 binding ELISA. BMS-378806 concentrations: ▪, 0 μM; ▴, 0.8 μM; ▾, 1.6 μM; ⧫, 3.2 μM. Data were analyzed according to the one-site binding model with GraphPad Prism. OD450, optical density at 450 nm. (B) Apparent binding constants for sCD4 in the absence and presence of BMS-378806. The Kd and Bmax values were derived by using nonlinear regression analysis with GraphPad Prism. Values were the means ± standard errors of the means, representing two independent experiments. (C) Binding affinity of BMS-378806 to gp120 by SPA. [3H]BMS-378806 binding to the gp120 protein was measured by employing gp120-coated SPA beads. The amount of gp120-bound [3H]BMS-378806 was determined by using a Packard TopCount scintillation counter. The results from Scatchard analysis of binding isotherm are shown in the insert. The x axis represents bound (corrected), and the y axis indicates bound/free.

Article Snippet: The curve represents the best fit to a saturation binding isotherm determined by using GraphPad Prism.

Techniques: Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay

Journal:

Article Title: Biochemical and Genetic Characterizations of a Novel Human Immunodeficiency Virus Type 1 Inhibitor That Blocks gp120-CD4 Interactions

doi: 10.1128/JVI.77.19.10528-10536.2003

Figure Lengend Snippet: Binding stoichiometry of BMS-378806 to gp120 protein. gp120JRFL (1.0 μM) was titrated with BMS-378806, and the observed percent reduction in fluorescence from each of four independent experiments was plotted first as a function of BMS-378806 concentration (data not shown). Each of these data sets was then normalized to the percent maximal fluorescence reduction observed for each experiment in order to analyze the four sets of data together. The individual, normalized data sets are depicted as open squares, closed squares, closed diamonds, and open triangles. The curve represents the best fit to a saturation binding isotherm determined by using GraphPad Prism.

Article Snippet: The curve represents the best fit to a saturation binding isotherm determined by using GraphPad Prism.

Techniques: Binding Assay, Fluorescence, Concentration Assay

Journal: Biochemical pharmacology

Article Title: Metabolism of bupropion by baboon hepatic and placental microsomes

doi: 10.1016/j.bcp.2011.04.014

Figure Lengend Snippet: A representative saturation curve of OH-BUP formation by baboon hepatic microsomes. The rate of bupropion hydroxylation to OH-BUP was dependent on bupropion concentration and exhibited typical saturation kinetics.

Article Snippet: The V max and apparent K m values were determined using nonlinear regression analysis of the saturation equation (GraphPad Prism 5, Vision 5.01, Graph Pad Software, Inc.).

Techniques: Concentration Assay

Journal: Biochemical pharmacology

Article Title: Metabolism of bupropion by baboon hepatic and placental microsomes

doi: 10.1016/j.bcp.2011.04.014

Figure Lengend Snippet: The representative saturation curve of threohydrobupropion formation by baboon placental microsomes (A) and the effect of chemical inhibitors selective to carbonyl reductases on the formation of threo- and erythrohydrobupropion by baboon placental microsomes (B). The inhibitors of carbonyl reductase are: 4-methylpyrazole (4-MP, 500µM), barbital (BAR, 500µM), flufenamic acid (FLU, 5µM), dicumarol (DIC, 500µM), menadione (MEN, 100µM), and 18β-glycyrrhetinic acid (18β-GA, 0.1µM). The rates of threo-(TB) and erythrohydrobupropion (EB) are expressed as percent of control (absence of inhibitor) and represent the mean ± S.D. of triplicate experiments. ** Statistical significance of p < 0.01 as determined by one-way ANOVA with Tukey’s comparison.

Article Snippet: The V max and apparent K m values were determined using nonlinear regression analysis of the saturation equation (GraphPad Prism 5, Vision 5.01, Graph Pad Software, Inc.).

Techniques: Control, Comparison