Coproteins [1]. Binding of gp120 to the receptor, CD4, on the target cell surface induces major conformational changes in the envelope glycoproteins [2]. These changes allow gp120 to bind the viral coreceptor, either CXCR4 or CCR5 [3?]. CD4 binding also induces the formation of a gp41 pre-hairpin intermediate, in which three hydrophobic grooves on the surface of a coiled coil formed by the heptad
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Tagonist `Compound A' [4-nitrobenzyl 1-(3(N-methylphenylsulfonamido)-3-phenylbutyl)piperidin-4-yl(vinyl)carbamate] [41,42] was kindly provided by Dr. Martin Springer at Merck Research Laboratories, Rahway, NJ. The anti-gp120 monoclonal antibody 48d, which recognizes an epitope that overlaps the coreceptor-binding site, was kindly provided by Dr. James Robinson (Tulane University Medical Center) [4
E sCD4 concentrations that are required to elicit shedding are significantly higher than those required to neutralize the virus [25,26]. In addition, for some HIV-1 strains, the temperature dependence of sCD4-induced gp120 shedding and virus neutralization differs [26]. The mode of sCD4-mediated inhibition thus remains incompletely understood. Targeting the functionally important and therefore con
Ues for the amount of antibody bound were obtained by subtracting binding measured in the absence of sCD4 from the binding measured at each time point after the sCD4 pulse.buffer, cells were incubated at room temperature for 45 minutes in FACS buffer containing a phycoerythrin-conjugated goat antihuman IgG antibody (Sigma) and an anti-CD4 fluoresceinconjugated antibody (OKT4, eBioscience) to measu
N the exposure of the coreceptor-binding site on gp120 and the HR1 coiled coil on gp41 after pulse activation by sCD4. To measure the progression of conformational changes that immediately follow HIV-1 envelope glycoprotein activation, we developed a novel enzyme-linked immunosorbent assay (ELISA)based system that utilizes live cells as a platform for expression of membrane-bound trimeric envelope
Ir Immune Defic Syndr. Author manuscript; available in PMC 2013 June 01.Mosam et al.Pagena e, 89 had advanced (but not immediately life threatening) KS and 58 had CD4