title: Adenosine-binding motif mimicry and cellular effects of a thieno[2,3-d]pyrimidine-based chemical inhibitor of atypical protein kinase C isoenzymes.
abstract: The aPKC [atypical PKC (protein kinase C)] isoforms ι and ζ play crucial roles in the formation and maintenance of cell polarity and represent attractive anti-oncogenic drug targets in Ras-dependent tumours. To date, few isoform-specific chemical biology tools are available to inhibit aPKC catalytic activity. In the present paper, we describe the identification and functional characterization of potent and selective thieno[2,3-d]pyrimidine-based chemical inhibitors of aPKCs. A crystal structure of human PKCι kinase domain bound to a representative compound, CRT0066854, reveals the basis for potent and selective chemical inhibition. Furthermore, CRT0066854 displaces a crucial Asn-Phe-Asp motif that is part of the adenosine-binding pocket and engages an acidic patch used by arginine-rich PKC substrates. We show that CRT0066854 inhibits the LLGL2 (lethal giant larvae 2) phosphorylation in cell lines and exhibits phenotypic effects in a range of cell-based assays. We conclude that this compound can be used as a chemical tool to modulate aPKC activity in vitro and in vivo and may guide the search for further aPKC-selective inhibitors.
authors: Structural Biology, Cancer Research UK, 44 Lincoln's Inn Fields, London WC2A 3LY, UK.
authors: Svend
authors: S
authors: Kjær
authors: Mark
authors: M
authors: Linch
authors: Andrew
authors: A
authors: Purkiss
authors: Brenda
authors: B
authors: Kostelecky
authors: Phillip P
authors: PP
authors: Knowles
authors: Carine
authors: C
authors: Rosse
authors: Philippe
authors: P
authors: Riou
authors: Christelle
authors: C
authors: Soudy
authors: Sarah
authors: S
authors: Kaye
authors: Bhavisha
authors: B
authors: Patel
authors: Erika
authors: E
authors: Soriano
authors: Judith
authors: J
authors: Murray-Rust
authors: Caroline
authors: C
authors: Barton
authors: Christian
authors: C
authors: Dillon
authors: Jon
authors: J
authors: Roffey
authors: Peter J
authors: PJ
authors: Parker
authors: Neil Q
authors: NQ
authors: McDonald
mesh_terms: D000241:Adenosine / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D018360:Crystallography, X-Ray; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004285:Dogs; D004353:Drug Evaluation, Preclinical / Q000379:methods; D057166:High-Throughput Screening Assays; D006801:Humans; D020128:Inhibitory Concentration 50; D007527:Isoenzymes / Q000037:antagonists & inhibitors; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D011743:Pyrimidines / Q000737:chemistry* / Q000494:pharmacology; D013876:Thiophenes / Q000737:chemistry / Q000494:pharmacology*
chemical_list: C581221:CRT0066854; D003598:Cytoskeletal Proteins; D007527:Isoenzymes; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D013876:Thiophenes; C526122:llgl2 protein, human; D000255:Adenosine Triphosphate; C115015:PKC-3 protein; D011493:Protein Kinase C; D000241:Adenosine