Download or read book Conformational Change Within Full-length HIV-1 Gp120 Upon Ligation with T-cell Receptor CD4 and Neutralizing Antibody IgG1 B12 written by Christopher Daniel Boone and published by . This book was released on 2009 with total page 164 pages. Available in PDF, EPUB and Kindle. Book excerpt: Viral membrane glycoprotein gpl20 is the key surface coat protein involved in HIV-1 human cellular recognition and initiating viral entry into the host cell. This viral entry is initiated by binding of gpl20 to the first ectodomain (Dl) of the hosts four domain T-cell receptor CD4. After binding with CD4, the variable V3 loop of gpl20 is extended out to ligate with a coreceptor on the T-cell surface, CXCR4 or CCR5. Small-angle X-ray scattering (SAXS)-derived structural perimeters and models of the unliganded soluble CD4 constructs (sCD4Di-D2, sCD4Di_D4) agreed well with previously reported crystallographic data with SCD4DI-D4 adapting a Z-conformation in solution. Likewise, SAXS data analyses of HIV-1 SFI62 gpl20 ligated with both sCD4 constructs revealed a conformational change in the D2-D3 linker region within SCD4DI-D4 which rotates the D3/D4 subdomains with respect to D1/D2 subdomains, changing the overall shape of SCD4DI-D4 from a Z- to a U-configuration upon binding gpl20. This conformational change of SCD4DI-D4 upon binding HIV-1 gpl20 is believed to be the key event that brings the extended V3 loop of gpl20 into close enough proximity with the T-cell coreceptor which then initiates viral entry into the host cell. Neutralizing antibody IgGl bl2 has been shown to clear a wide array of HIV-1 strains by binding to gpl20 at the CD4 epitope. The crystal structure of this antibody displays a highly asymmetric conformation with a shift of the Fc domain so that it lies directly underneath one of the Fab domains (FabC) while the other Fab domain is extended out (FabE). Our SAXS-derived models for unliganded IgGl bl2 revealed that this characteristic asymmetric shape is the predominate conformation in solution where the antibody is free of crystal packing forces. Upon mixing a 1:2 molar ratio of IgGl bl2 and HIV-leaL gpl20, SAXS analyses indicate that the predominate scattering entity is the 1:1 antibody/antigen binary complex and not the expected 1:2 Ab/Ag ternary structure. Placement of known crystallographic data within our SAXS-based models for the 1:1 IgGl bl2/HIV-lBaL gpl20 binary complex suggests that only the FabE domain binds to gpl20 while leaving very other little conformational change to the asymmetric profile of the antibody as well as in the viral glycoprotein. This 1:1 binary complex may be the result of steric hindrance at the two Fab binding sites created via ligation of two antigen particles. Unusual rigidity encoded in the core and lower hinge regions of IgGl bl2 may construct the characteristic asymmetric shape of the antibody as well as promoting 1:1 binding with HIV-leaL gpl20. Small-angle neutron scattering (SANS) of unliganded HIV-1SFI62 gpl20 and the gpl20/sCD4Di-D2 binary complex was collected at various solvent D2O levels. This contrast series was carried out to reveal possible conformational changes of the extensive surface glycoslyation of HIV-1SFI62 gpl20 upon binding sCD4Di. D2- SANS-derived models and analysis of structural parameters revealed a possible candidate for the scattering density of the V1/V2 emanated loop of HIV-1SFI62 gpl20 known to coordinate ligation of CD4 as well as a possible shift in the surface glycosylation of gpl20 upon binding SCD4DI-D2.