title: HIV gp41 six-helix bundle constructs induce rapid vesicle fusion at pH 3.5 and little fusion at pH 7.0: understanding pH dependence of protein aggregation, membrane binding, and electrostatics, and implications for HIV-host cell fusion.
abstract: The HIV gp41 protein catalyzes fusion between HIV and target cell membranes. The fusion states of the gp41 ectodomain include early coiled-coil (CC) structure and final six-helix bundle (SHB) structure. The ectodomain has an additional N-terminal apolar fusion peptide (FP) sequence which binds to target cell membranes and plays a critical role in fusion. One approach to understanding gp41 function is study of vesicle fusion induced by constructs that encompass various regions of gp41. There are apparent conflicting literature reports of either rapid or no fusion of negatively charged vesicles by SHB constructs. These reports motivated the present study, which particularly focused on effects of pH because the earlier high and no fusion results were at pH 3.0 and 7.2, respectively. Constructs include "Hairpin," which has SHB structure but lacks the FP, "FP-Hairpin" with FP + SHB, and "N70," which contains the FP and part of the CC but does not have SHB structure. Aqueous solubility, membrane binding, and vesicle fusion function were measured at a series of pHs and much of the pH dependences of these properties were explained by protein charge. At pH 3.5, all constructs were positively charged, bound negatively charged vesicles, and induced rapid fusion. At pH 7.0, N70 remained positively charged and induced rapid fusion, whereas Hairpin and FP-Hairpin were negatively charged and induced no fusion. Because viral entry occurs near pH 7 rather than pH 3, our results are consistent with fusogenic function of early CC gp41 and with fusion arrest by final SHB gp41.
authors.affiliation: Department of Chemistry, Michigan State University, East Lansing, MI 48824, USA.
authors.forename: Kelly
authors.initials: K
authors.lastname: Sackett
authors.forename: Allan
authors.initials: A
authors.lastname: TerBush
authors.forename: David P
authors.initials: DP
authors.lastname: Weliky
mesh_terms: D000595:Amino Acid Sequence; D002459:Cell Fusion; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D006678:HIV / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D015700:HIV Envelope Protein gp41 / Q000737:chemistry / Q000494:pharmacology*; D015658:HIV Infections / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion / Q000187:drug effects*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000187:drug effects; D017510:Protein Folding / Q000187:drug effects; D017433:Protein Structure, Secondary; D012995:Solubility; D012996:Solutions / Q000737:chemistry; D055672:Static Electricity
chemical_list: D015700:HIV Envelope Protein gp41; D012996:Solutions