title: Expression and purification of myristoylated matrix protein of Mason-Pfizer monkey virus for NMR and MS measurements.
abstract: Matrix proteins play multiple roles both in early and late stages of the viral replication cycle. Their N-terminal myristoylation is important for interaction with the host cell membrane during virus budding. We used Escherichia coli, carrying N-myristoyltransferase gene, for the expression of the myristoylated His-tagged matrix protein of Mason-Pfizer monkey virus. An efficient, single-step purification procedure eliminating all contaminating proteins including, importantly, the non-myristoylated matrix protein was designed. The comparison of NMR spectra of matrix protein with its myristoylated form revealed substantial structural changes induced by this fatty acid modification.
authors.affiliation: Laboratory of NMR Spectroscopy, Institute of Chemical Technology, Prague, Czech Republic.
authors.forename: Jan
authors.initials: J
authors.lastname: Prchal
authors.forename: Petra
authors.initials: P
authors.lastname: Junkova
authors.forename: Miroslava
authors.initials: M
authors.lastname: Strmiskova
authors.forename: Jan
authors.initials: J
authors.lastname: Lipov
authors.forename: Radovan
authors.initials: R
authors.lastname: Hynek
authors.forename: Tomas
authors.initials: T
authors.lastname: Ruml
authors.forename: Richard
authors.initials: R
authors.lastname: Hrabal
mesh_terms: D000217:Acyltransferases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D015870:Gene Expression; D016093:Mason-Pfizer monkey virus / Q000737:chemistry / Q000235:genetics*; D019814:Myristic Acid / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014763:Viral Matrix Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D019814:Myristic Acid; D000217:Acyltransferases; C050480:glycylpeptide N-tetradecanoyltransferase