title: Structural basis for phosphorylation and lysine acetylation cross-talk in a kinase motif associated with myocardial ischemia and cardioprotection.
abstract: Myocardial ischemia and cardioprotection by ischemic pre-conditioning induce signal networks aimed at survival or cell death if the ischemic period is prolonged. These pathways are mediated by protein post-translational modifications that are hypothesized to cross-talk with and regulate each other. Phosphopeptides and lysine-acetylated peptides were quantified in isolated rat hearts subjected to ischemia or ischemic pre-conditioning, with and without splitomicin inhibition of lysine deacetylation. We show lysine acetylation (acetyl-Lys)-dependent activation of AMP-activated protein kinase, AKT, and PKA kinases during ischemia. Phosphorylation and acetyl-Lys sites mapped onto tertiary structures were proximal in >50% of proteins investigated, yet they were mutually exclusive in 50 ischemic pre-conditioning- and/or ischemia-associated peptides containing the KXXS basophilic protein kinase consensus motif. Modifications in this motif were modeled in the C terminus of muscle-type creatine kinase. Acetyl-Lys increased proximal dephosphorylation by 10-fold. Structural analysis of modified muscle-type creatine kinase peptide variants by two-dimensional NMR revealed stabilization via a lysine-phosphate salt bridge, which was disrupted by acetyl-Lys resulting in backbone flexibility and increased phosphatase accessibility.
authors.affiliation: From the Discipline of Pathology, School of Medical Sciences, University of Sydney, Sydney 2006, Australia, the Diabetes and Obesity Program, Biological Mass Spectrometry Unit, Garvan Institute of Medical Research, 2010 Australia.
authors.forename: Benjamin L
authors.initials: BL
authors.lastname: Parker
authors.affiliation: the School of Molecular Bioscience and.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Shepherd
authors.affiliation: the Diabetes and Obesity Program.
authors.forename: Sophie
authors.initials: S
authors.lastname: Trefely
authors.affiliation: the Diabetes and Obesity Program, Biological Mass Spectrometry Unit, Garvan Institute of Medical Research, 2010 Australia.
authors.forename: Nolan J
authors.initials: NJ
authors.lastname: Hoffman
authors.affiliation: the School of Molecular Bioscience and the Charles Perkins Centre, University of Sydney, Sydney 2006, Australia, and.
authors.forename: Melanie Y
authors.initials: MY
authors.lastname: White
authors.affiliation: the Charles Perkins Centre, University of Sydney, Sydney 2006, Australia, and.
authors.forename: Kasper
authors.initials: K
authors.lastname: Engholm-Keller
authors.affiliation: From the Discipline of Pathology, School of Medical Sciences, University of Sydney, Sydney 2006, Australia, the Charles Perkins Centre, University of Sydney, Sydney 2006, Australia, and.
authors.forename: Brett D
authors.initials: BD
authors.lastname: Hambly
authors.affiliation: the Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark.
authors.forename: Martin R
authors.initials: MR
authors.lastname: Larsen
authors.affiliation: the Diabetes and Obesity Program, Biological Mass Spectrometry Unit, Garvan Institute of Medical Research, 2010 Australia, the Charles Perkins Centre, University of Sydney, Sydney 2006, Australia, and.
authors.forename: David E
authors.initials: DE
authors.lastname: James
authors.affiliation: From the Discipline of Pathology, School of Medical Sciences, University of Sydney, Sydney 2006, Australia, the School of Molecular Bioscience and the Charles Perkins Centre, University of Sydney, Sydney 2006, Australia, and stuart.cordwell@sydney.edu.au.
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Cordwell
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000096:biosynthesis; D000107:Acetylation / Q000187:drug effects; D020816:Amino Acid Motifs; D000818:Animals; D002316:Cardiotonic Agents / Q000008:administration & dosage; D019194:Ischemic Preconditioning; D008239:Lysine / Q000378:metabolism*; D017202:Myocardial Ischemia / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D009281:Naphthalenes / Q000008:administration & dosage; D010766:Phosphorylation; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D051057:Proto-Oncogene Proteins c-akt / Q000096:biosynthesis; D011753:Pyrones / Q000008:administration & dosage; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics
chemical_list: D002316:Cardiotonic Agents; D009281:Naphthalenes; D011753:Pyrones; C444426:splitomicin; D010770:Phosphotransferases; D051057:Proto-Oncogene Proteins c-akt; D055372:AMP-Activated Protein Kinases; D008239:Lysine
keywords: Cardioprotection; Heart; Lysine Acetylation; Mass Spectrometry (MS); Myocardial Infarction; Phosphorylation; Post-translational Modification (PTM); Proteomics; Signaling