HIV fusion peptide and its cross-linked oligomers: efficient syntheses, significance of the trimer in fusion activity, correlation of beta strand conformation with membrane cholesterol, and proximity to lipid headgroups.
For enveloped viruses such as HIV, an approximately 20-residue N-terminal fusion peptide domain in the envelope protein binds to target cell membranes and plays a key role in fusion between the viral and cellular membranes during infection. The chemically synthesized HIV fusion peptide (HFP) catalyzes fusion between membrane vesicles and is a useful model system for understanding some aspects of HIV fusion. Previous studies have shown a common trimeric state for the envelope protein from several different viruses, including HIV, and in this study, practical high-yield syntheses are reported for HFP monomer (HFPmn) and chemically cross-linked HFP dimer (HFPdm), trimer (HFPtr), and tetramer (HFPte). The vesicle fusion rates per strand were ordered as follows: HFPmn < HFPdm < HFPtr approximately HFPte. This suggested that HFPtr is the smallest catalytically efficient oligomer. Solid-state NMR measurements of (13)CO chemical shifts were carried out in constructs labeled at either Ala-6 or Ala-15. For all constructs associated with cholesterol-containing membranes, the chemical shifts of both residues correlated with beta strand conformation while association with membranes without cholesterol resulted in a mixture of helical and beta strand conformations. The dependence of fusion rate on oligomer size is independent of membrane cholesterol content, so one interpretation of the data is fusion activity of both helical and beta strand conformations. Membrane location may be a determinant of fusion activity, and for all constructs in both conformations, a large fraction of the Ala-15 (13)CO groups were 5-6 A from the (31)P atoms in the lipid headgroups, while the Ala-6 (13)CO groups were more distant.
Qiang
Wei
W
Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
Weliky
David P
DP
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry* / Q000378:metabolism; D003198:Computer Simulation; D003432:Cross-Linking Reagents / Q000737:chemistry; D006678:HIV / Q000378:metabolism*; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008055:Lipids / Q000737:chemistry*; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013696:Temperature
D003432:Cross-Linking Reagents; D015700:HIV Envelope Protein gp41; D008055:Lipids; D010455:Peptides; D002784:Cholesterol