title: Conformational properties of peptides corresponding to the ebolavirus GP2 membrane-proximal external region in the presence of micelle-forming surfactants and lipids.
abstract: Ebola virus and Sudan virus are members of the family Filoviridae of nonsegmented negative-strand RNA viruses ("filoviruses") that cause severe hemorrhagic fever with fatality rates as high as 90%. Infection by filoviruses requires membrane fusion between the host and the virus; this process is facilitated by the two subunits of the envelope glycoprotein, GP1 (the surface subunit) and GP2 (the transmembrane subunit). The membrane-proximal external region (MPER) is a Trp-rich segment that immediately precedes the transmembrane domain of GP2. In the analogous glycoprotein for HIV-1 (gp41), the MPER is critical for membrane fusion and is the target of several neutralizing antibodies. However, the role of the MPER in filovirus GP2 and its importance in membrane fusion have not been established. Here, we characterize the conformational properties of peptides representing the GP MPER segments of Ebola virus and Sudan virus in the presence of micelle-forming surfactants and lipids, at pH 7 and 4.6. Circular dichroism spectroscopy and tryptophan fluorescence indicate that the GP2 MPER peptides bind to micelles of sodium dodecyl sulfate and dodecylphosphocholine (DPC). Nuclear magnetic resonance spectroscopy of the Sudan virus MPER peptide revealed that residues 644-651 interact directly with DPC, and that this interaction enhances the helical conformation of the peptide. The Sudan virus MPER peptide was found to moderately inhibit cell entry by a GP-pseudotyped vesicular stomatitis virus but did not induce leakage of a fluorescent molecule from a large unilammellar vesicle comprised of 1-palmitoyl-2-oleoylphosphatidylcholine or cause hemolysis. Taken together, this analysis suggests the filovirus GP2 MPER binds and inserts shallowly into lipid membranes.
authors: Department of Biochemistry and ‡Department of Microbiology and Immunology, Albert Einstein College of Medicine , 1300 Morris Park Avenue, Bronx, New York 10461, United States.
authors: Lauren K
authors: LK
authors: Regula
authors: Richard
authors: R
authors: Harris
authors: Fang
authors: F
authors: Wang
authors: Chelsea D
authors: CD
authors: Higgins
authors: Jayne F
authors: JF
authors: Koellhoffer
authors: Yue
authors: Y
authors: Zhao
authors: Kartik
authors: K
authors: Chandran
authors: Jianmin
authors: J
authors: Gao
authors: Mark E
authors: ME
authors: Girvin
authors: Jonathan R
authors: JR
authors: Lai
mesh_terms: D000595:Amino Acid Sequence; D029043:Ebolavirus / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008823:Micelles; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D013501:Surface-Active Agents / Q000737:chemistry / Q000378:metabolism; D014364:Tryptophan / Q000235:genetics / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D008823:Micelles; D010455:Peptides; D013501:Surface-Active Agents; D014759:Viral Envelope Proteins; C113189:envelope glycoprotein, Ebola virus; D010767:Phosphorylcholine; C028810:dodecylphosphocholine; D014364:Tryptophan
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