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- W2045176853 abstract "The HIV-1 envelope (Env) transmembrane protein, gp41, is typically considered a type I membrane protein with an extracellular N-terminus, a single membrane-spanning domain, and a C-terminus forming a ∼150 residue intracytoplasmic tail. However, published studies indicate an alternative or dynamic topology for portions of the C-terminal tail (CTT) that results in exposure of CTT segments on the membrane surface. To distinguish between these alternative models, we evaluated the accessibility of a reference CTT sequence, the “Kennedy epitope” (KE), in viral and cellular membranes to map CTT topology relative to the lipid bilayer. KE accessibility in cell-associated Env was defined by reactivity of native or VSV-G epitope-tagged KE to specific monoclonal antibodies (MAbs), measured by FACS of intact cells. In parallel, KE exposure in virion-associated Env was characterized by MAb binding to intact virions as measured in immunoprecipitation and surface plasmon resonance (SPR) spectroscopy assays. FACS analyses of live cell-associated Env demonstrated significant reactivity of the KE with MAbs to native or VSV-G epitope-tagged KE. No reactivity was observed in cells expressing Env with the VSV-G epitope substituted into a cytoplasmic domain. In contrast to cellular Env, KE in the context of virions failed to react with MAbs directed to the native KE sequence, as measured both by immunoprecipitation and SPR assays. However, MAbs specific for a membrane-proximal sequence bound virions in both assays. Together, the results of these accessibility assays indicate that the KE sequences of gp41 are accessible to antibody binding in cell surface-expressed, but not virion-associated, Env. These observations suggest that the CTT may assume distinct topologies (reflected in KE exposure) that depend on the membrane environment (viral/cellular) and that parts of the CTT may be (transiently) exposed on the membrane surface." @default.
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- W2045176853 date "2010-01-01" @default.
- W2045176853 modified "2023-09-26" @default.
- W2045176853 title "Distinct Topologies For the HIV-1 Transmembrane Glycoprotein gp41 C-Terminal Tail on Cellular and Viral Lipid Membranes" @default.
- W2045176853 doi "https://doi.org/10.1016/j.bpj.2009.12.282" @default.
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