Structural and Thermodynamic Analysis of HIV-1 Fusion Inhibition Using Small gp41 Mimetic Proteins. Issue 17 (9th August 2019)

• Record Type: Journal Article
• Authors: Jurado, Samuel; Cano-Muñoz, Mario; Morel, Bertrand; Standoli, Sara; Santarossa, Elisabetta; Moog, Christiane; Schmidt, Sylvie; Laumond, Géraline; Cámara-Artigas, Ana; Conejero-Lara, Francisco
• Is Part Of: Journal of molecular biology. Volume 431:Issue 17(2019)
• Page Start: 3091
• Page End: 3106
• Publication Date: 2019-08-09
• Subjects: Env HIV-1 envelope glycoprotein -- gp41 glycoprotein subunit 41 -- gp120 glycoprotein subunit 120 -- CD circular dichroism spectroscopy -- ITC isothermal titration calorimetry -- DLS dynamic light scattering -- DSC differential scanning calorimetry -- SASA solvent accessible surface area; envelope glycoprotein -- fusion inhibitor -- calorimetry -- binding -- x-ray crystallography; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.06.022
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: Development of effective inhibitors of the fusion between HIV-1 and the host cell membrane mediated by gp41 continues to be a grand challenge due to an incomplete understanding of the molecular and mechanistic details of the fusion process. We previously developed single-chain, chimeric proteins (named covNHR) that accurately mimic the N-heptad repeat (NHR) region of gp41 in a highly stable coiled-coil conformation. These molecules bind strongly to peptides derived from the gp41 C-heptad repeat (CHR) and are potent and broad HIV-1 inhibitors. Here, we investigated two covNHR variants differing in two mutations, V10E and Q123R (equivalent to V38E and Q40R in gp41 sequence) that reproduce the effect of HIV-1 mutations associated with resistance to fusion inhibitors, such as T20 (enfuvirtide). A detailed calorimetric analysis of the binding between the covNHR proteins and CHR peptides (C34 and T20) reveals drastic changes in affinity due to the mutations as a result of local changes in interactions at the site of T20 resistance. The crystallographic structure of the covNHR:C34 complex shows a virtually identical CHR–NHR binding interface to that of the post-fusion structure of gp41 and underlines an important role of buried interfacial water molecules in binding affinity and in development of resistance against CHR peptides. Despite the great difference in affinity, both covNHR variants demonstrate strong inhibitory activity for a wide variety of HIV-1 strains. These properties support the high potential of these covNHR proteins as new potent HIV-1 inhibitors. Our results may guide future inhibition approaches. Graphical Abstract: Unlabelled Image Highlights: CovNHR proteins are good mimics of HIV-1 gp41 coiled-coil. X-ray structure of the complex between covNHR and C34 peptide was solved. Two mutations related to enfuvirtide resistance strongly reduce affinity for C34. Interfacial water molecules mediate high CHR-NHR binding affinity. Enfuvirtide-resistance mutations have a small effect on covNHR inhibitory activity.