title: New conformational properties induced by the replacement of Tyr-64 in Desulfovibrio vulgaris Hildenborough ferricytochrome c553 using isotopic exchanges monitored by mass spectrometry.
abstract: In order to study the conformational stability induced by the replacement of Tyr-64 in Desulfovibrio vulgaris Hildenborough (DvH) cytochrome c553, fast peptic digestion of deuterated protein followed by separation and measurement of related peptides using liquid chromatography coupled to electrospray ionization mass spectrometry was performed. We show that the H-bonding and/or solvent accessibility properties were modified by the single-site mutation. The mutant proteins can be classified into two groups: the Y64F and Y64L mutants with nearly unchanged deuterium incorporation compared to the wild-type protein and the Y64S, Y64V and Y64A mutants with increased deuterium incorporation. The 70-74 peptide was the most affected by mutation of Tyr-64, the phenylalanine mutant inducing slight stabilization whereas the serine mutant was significantly destabilized. In addition, from the analysis of the overlapping 37-57 and 38-57 peptides we can conclude that the amide proton of Tyr-38 has been replaced by deuterium in all proteins.
authors.affiliation: Institut de Biologie Structurale Jean-Pierre Ebel, Grenoble, France.
authors.forename: P
authors.initials: P
authors.lastname: Guy
authors.forename: M
authors.initials: M
authors.lastname: Jaquinod
authors.forename: H
authors.initials: H
authors.lastname: Rémigy
authors.forename: J P
authors.initials: JP
authors.lastname: Andrieu
authors.forename: J
authors.initials: J
authors.lastname: Gagnon
authors.forename: B
authors.initials: B
authors.lastname: Bersch
authors.forename: A
authors.initials: A
authors.lastname: Dolla
authors.forename: L
authors.initials: L
authors.lastname: Blanchard
authors.forename: F
authors.initials: F
authors.lastname: Guerlesquin
authors.forename: E
authors.initials: E
authors.lastname: Forest
mesh_terms: D003574:Cytochrome c Group / Q000737:chemistry*; D016969:Desulfovibrio vulgaris / Q000737:chemistry*; D003903:Deuterium; D004583:Electrons; D006860:Hydrogen Bonding; D008970:Molecular Weight; D009154:Mutation; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011487:Protein Conformation; D012997:Solvents; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014443:Tyrosine / Q000502:physiology*
chemical_list: D003574:Cytochrome c Group; D010446:Peptide Fragments; D012997:Solvents; C083951:cytochrome c553; D014443:Tyrosine; D003903:Deuterium