title: Selective Phosphorylation Inhibitor of Delta Protein Kinase C-Pyruvate Dehydrogenase Kinase Protein-Protein Interactions: Application for Myocardial Injury in Vivo.
abstract: Protein kinases regulate numerous cellular processes, including cell growth, metabolism, and cell death. Because the primary sequence and the three-dimensional structure of many kinases are highly similar, the development of selective inhibitors for only one kinase is challenging. Furthermore, many protein kinases are pleiotropic, mediating diverse and sometimes even opposing functions by phosphorylating multiple protein substrates. Here, we set out to develop an inhibitor of a selective protein kinase phosphorylation of only one of its substrates. Focusing on the pleiotropic delta protein kinase C (δPKC), we used a rational approach to identify a distal docking site on δPKC for its substrate, pyruvate dehydrogenase kinase (PDK). We reasoned that an inhibitor of PDK's docking should selectively inhibit the phosphorylation of only PDK without affecting phosphorylation of the other δPKC substrates. Our approach identified a selective inhibitor of PDK docking to δPKC with an in vitro Kd of ∼50 nM and reducing cardiac injury IC50 of ∼5 nM. This inhibitor, which did not affect the phosphorylation of other δPKC substrates even at 1 μM, demonstrated that PDK phosphorylation alone is critical for δPKC-mediated injury by heart attack. The approach we describe is likely applicable for the identification of other substrate-specific kinase inhibitors.
authors.affiliation: Department of Chemical and Systems Biology, School of Medicine, Stanford University , Stanford, California 94305-5174, United States.
authors.forename: Nir
authors.initials: N
authors.lastname: Qvit
authors.affiliation: Department of Chemical and Systems Biology, School of Medicine, Stanford University , Stanford, California 94305-5174, United States.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Disatnik
authors.affiliation: Nanjing, Jiangsu 210000, P.R. China.
authors.forename: Eiketsu
authors.initials: E
authors.lastname: Sho
authors.affiliation: Department of Chemical and Systems Biology, School of Medicine, Stanford University , Stanford, California 94305-5174, United States.
authors.forename: Daria
authors.initials: D
authors.lastname: Mochly-Rosen
mesh_terms: D000818:Animals; D008297:Male; D008807:Mice, Inbred BALB C; D062105:Molecular Docking Simulation; D015428:Myocardial Reperfusion Injury / Q000188:drug therapy* / Q000201:enzymology; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000627:therapeutic use / Q000633:toxicity; D010766:Phosphorylation; D011485:Protein Binding; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000627:therapeutic use / Q000633:toxicity; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D011994:Recombinant Proteins; D013379:Substrate Specificity
chemical_list: D010455:Peptides; D047428:Protein Kinase Inhibitors; D000081382:Pyruvate Dehydrogenase Acetyl-Transferring Kinase; D011994:Recombinant Proteins; D017346:Protein Serine-Threonine Kinases; D051745:Protein Kinase C-delta