Cell Surface Assembly of HIV gp41 Six-Helix Bundles for Facile, Quantitative Measurements of Hetero-oligomeric Interactions

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• 作者：Xuebo Hu ; Piyali Saha ; Xiaoyue Chen ; Dogeun Kim ; Mahesh Devarasetty ; Raghavan Varadarajan ; Moonsoo M. Jin
• 刊名：The Journal of the American Chemical Society
• 出版年：2012
• 出版时间：September 12, 2012
• 年：2012
• 卷：134
• 期：36
• 页码：14642-14645
• 全文大小：333K
• 年卷期：v.134,no.36(September 12, 2012)
• ISSN：1520-5126

文摘

Helix鈥揾elix interactions are fundamental to many biological signals and systems and are found in homo- or heteromultimerization of signaling molecules as well as in the process of virus entry into the host. In HIV, virus鈥揾ost membrane fusion during infection is mediated by the formation of six-helix bundles (6HBs) from homotrimers of gp41, from which a number of synthetic peptides have been derived as antagonists of virus entry. Using a yeast surface two-hybrid (YS2H) system, a platform designed to detect protein鈥損rotein interactions occurring through a secretory pathway, we reconstituted 6HB complexes on the yeast surface, quantitatively measured the equilibrium and kinetic constants of soluble 6HB, and delineated the residues influencing homo-oligomeric and hetero-oligomeric coiled-coil interactions. Hence, we present YS2H as a platform for the facile characterization and design of antagonistic peptides for inhibition of HIV and many other enveloped viruses relying on membrane fusion for infection, as well as cellular signaling events triggered by hetero-oligomeric coiled coils.