title: Membrane Interactions of the Mason-Pfizer Monkey Virus Matrix Protein and Its Budding Deficient Mutants.
abstract: Matrix proteins (MAs) play a key role in the transport of retroviral proteins inside infected cells and in the interaction with cellular membranes. In most retroviruses, retroviral MAs are N-terminally myristoylated. This modification serves as a membrane targeting signal and also as an anchor for membrane interaction. The aim of this work was to characterize the interactions anchoring retroviral MA at the plasma membrane of infected cell. To address this issue, we compared the structures and membrane affinity of the Mason-Pfizer monkey virus (M-PMV) wild-type MA with its two budding deficient double mutants, that is, T41I/T78I and Y28F/Y67F. The structures of the mutants were determined using solution NMR spectroscopy, and their interactions with water-soluble phospholipids were studied. Water-soluble phospholipids are widely used models for studying membrane interactions by solution NMR spectroscopy. However, this approach might lead to artificial results due to unnatural hydrophobic interactions. Therefore, we used a new approach based on the measurement of the loss of the1H NMR signal intensity of the protein sample induced by the addition of the liposomes containing phospholipids with naturally long fatty acids. HIV-1 MA was used as a positive control because its ability to interact with liposomes has already been described. We found that in contrast to HIV-1, the M-PMV MA interacted with the liposomes differently and much weaker. In our invivo experiments, the M-PMV MA did not co-localize with lipid rafts. Therefore, we concluded that M-PMV might adopt a different membrane binding mechanism than HIV-1.
authors.affiliation: Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic; Laboratory of NMR Spectroscopy, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: kroupat@vscht.cz.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Kroupa
authors.affiliation: Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: langeroh@vscht.cz.
authors.forename: Hana
authors.initials: H
authors.lastname: Langerová
authors.affiliation: Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic; Laboratory of NMR Spectroscopy, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: dolezal@uochb.cas.cz.
authors.forename: Michal
authors.initials: M
authors.lastname: Doležal
authors.affiliation: Laboratory of NMR Spectroscopy, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: prchalj@vscht.cz.
authors.forename: Jan
authors.initials: J
authors.lastname: Prchal
authors.affiliation: Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: spiwokv@vscht.cz.
authors.forename: Vojtěch
authors.initials: V
authors.lastname: Spiwok
authors.affiliation: Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road, Atlanta, GA 30329, USA. Electronic address: ehunte4@emory.edu.
authors.forename: Eric
authors.initials: E
authors.lastname: Hunter
authors.affiliation: Department of Biotechnology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: rumlovai@vscht.cz.
authors.forename: Michaela
authors.initials: M
authors.lastname: Rumlová
authors.affiliation: Laboratory of NMR Spectroscopy, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: hrabalr@vscht.cz.
authors.forename: Richard
authors.initials: R
authors.lastname: Hrabal
authors.affiliation: Department of Biochemistry and Microbiology, University of Chemistry and Technology, Prague, Technická 5, 166 28 Prague 6, Czech Republic. Electronic address: rumlt@vscht.cz.
authors.forename: Tomáš
authors.initials: T
authors.lastname: Ruml
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D016093:Mason-Pfizer monkey virus / Q000502:physiology*; D050505:Mutant Proteins / Q000737:chemistry* / Q000378:metabolism*; D020125:Mutation, Missense; D010743:Phospholipids / Q000378:metabolism; D011485:Protein Binding; D014763:Viral Matrix Proteins / Q000737:chemistry* / Q000378:metabolism*; D057074:Virus Release
chemical_list: D005227:Fatty Acids; D008081:Liposomes; D050505:Mutant Proteins; D010743:Phospholipids; D014763:Viral Matrix Proteins
keywords: Mason-Pfizer monkey virus; budding; liposome binding; matrix protein; membrane interaction