title: Characterization of metal and nucleotide liganded forms of adenylate kinase by electrospray ionization mass spectrometry.
abstract: Complexes of adenylate kinase from Escherichia coli, Bacillus subtilis, and Bacillus stearothermophilus with the bisubstrate nucleotide analog P1,P5-di(adenosine 5')-pentaphosphate and with metal ions (Zn2+ and/or Mg2+) were analyzed by electrospray ionization mass spectrometry. P1,P5-di(adenosine 5')-pentaphosphate. adenylate kinase complex was detected in the positive mode at pH as low as 3.8. Binding of nucleotide to adenylate kinase stabilizes the overall structure of the protein and preserves the Zn2+ chelated form of the enzyme from the gram-positive organisms. In this way, it is possible in a single mass spectrometry experiment to screen metal-chelating adenylate kinases, without use of radioactively labeled compounds. Binding of Mg2+ to enzyme via P1,P5-di(adenosine 5')-pentaphosphate was also demonstrated by mass spectrometry. Although no amino acid side chain in adenylate kinase is supposed to interact with Mg2+, Asp93 in porcine muscle cytosolic enzyme, equivalent to Asp84 in the E. coli adenylate kinase, was proposed to stabilize the nucleotide.Mg2+ complex via water molecules.
authors.affiliation: Laboratoire d'Application de Spectrométrie de Masse, Université de Lille II, Lille Cedex, 59045, France.
authors.forename: G
authors.initials: G
authors.lastname: Briand
authors.forename: V
authors.initials: V
authors.lastname: Perrier
authors.forename: M
authors.initials: M
authors.lastname: Kouach
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.forename: A M
authors.initials: AM
authors.lastname: Gilles
authors.forename: O
authors.initials: O
authors.lastname: Bârzu
mesh_terms: D000263:Adenylate Kinase / Q000737:chemistry* / Q000378:metabolism; D001412:Bacillus subtilis; D004926:Escherichia coli; D001411:Geobacillus stearothermophilus; D008024:Ligands; D013058:Mass Spectrometry; D008670:Metals / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D011485:Protein Binding
chemical_list: D008024:Ligands; D008670:Metals; D009711:Nucleotides; D000263:Adenylate Kinase