title: N-myristoylated c-Abl tyrosine kinase localizes to the endoplasmic reticulum upon binding to an allosteric inhibitor.
abstract: Allosteric kinase inhibitors hold promise for revealing unique features of kinases that may not be apparent using conventional ATP-competitive inhibitors. Here we explore the activity of a previously reported allosteric inhibitor of BCR-Abl kinase, GNF-2, against two cellular isoforms of Abl tyrosine kinase: one that carries a myristate in the N terminus and the other that is deficient in N-myristoylation. Our results show that GNF-2 inhibits the kinase activity of non-myristoylated c-Abl more potently than that of myristoylated c-Abl by binding to the myristate-binding pocket in the C-lobe of the kinase domain. Unexpectedly, indirect immunofluorescence reveals a translocation of myristoylated c-Abl to the endoplasmic reticulum in GNF-2-treated cells, whereas GNF-2 has no detectable effect on the localization of non-myristoylated c-Abl. These results indicate that GNF-2 competes with the NH(2)-terminal myristate for binding to the c-Abl kinase myristate-binding pocket and that the exposed myristoyl group accounts for the localization to the endoplasmic reticulum. We also demonstrate that GNF-2 can inhibit enzymatic and cellular kinase activity of Arg, a kinase highly homologous to c-Abl, which is also likely to be regulated through intramolecular binding of an NH(2)-terminal myristate lipid. These results suggest that non-ATP-competitive inhibitors, such as GNF-2, can serve as chemical tools that can discriminate between c-Abl isoform-specific behaviors.
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.
authors.forename: Yongmun
authors.initials: Y
authors.lastname: Choi
authors.forename: Markus A
authors.initials: MA
authors.lastname: Seeliger
authors.forename: Shoghag B
authors.initials: SB
authors.lastname: Panjarian
authors.forename: Hakjoong
authors.initials: H
authors.lastname: Kim
authors.forename: Xianming
authors.initials: X
authors.lastname: Deng
authors.forename: Taebo
authors.initials: T
authors.lastname: Sim
authors.forename: Brian
authors.initials: B
authors.lastname: Couch
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Koleske
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Smithgall
authors.forename: Nathanael S
authors.initials: NS
authors.lastname: Gray
mesh_terms: D016475:3T3 Cells; D000255:Adenosine Triphosphate / Q000737:chemistry; D000495:Allosteric Site; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001665:Binding Sites; D019008:Drug Resistance, Neoplasm; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D006801:Humans; D008055:Lipids / Q000737:chemistry; D051379:Mice; D008968:Molecular Conformation; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D016315:Proto-Oncogene Proteins c-abl / Q000378:metabolism / Q000502:physiology*
chemical_list: D000970:Antineoplastic Agents; D008055:Lipids; D020033:Protein Isoforms; D000255:Adenosine Triphosphate; D016315:Proto-Oncogene Proteins c-abl
references.citation: Structure. 2005 Jun;13(6):861-71
references.pmid: 15939018
references.citation: Nature. 1995 Feb 9;373(6514):536-9
references.pmid: 7845468
references.citation: J Cell Biol. 1998 May 18;141(4):887-94
references.pmid: 9585408
references.citation: Genes Dev. 1997 Oct 1;11(19):2456-67
references.pmid: 9334312
references.citation: J Mol Biol. 2008 Jan 4;375(1):70-9
references.pmid: 17999933
references.citation: Oncogene. 2008 Jul 24;27(32):4385-91
references.pmid: 18391983
references.citation: Nat Biotechnol. 2008 Jan;26(1):127-32
references.pmid: 18183025
references.citation: Nat Chem Biol. 2008 Aug;4(8):474-82
references.pmid: 18641628
references.citation: Neuron. 1998 Dec;21(6):1259-72
references.pmid: 9883720
references.citation: Proc Natl Acad Sci U S A. 2004 Dec 14;101(50):17456-61
references.pmid: 15505216
references.citation: Nat Cell Biol. 2004 Jan;6(1):3-7
references.pmid: 14704671
references.citation: Cell. 2003 Mar 21;112(6):859-71
references.pmid: 12654251
references.citation: J Biol Chem. 2001 Jul 20;276(29):27104-10
references.pmid: 11309382
references.citation: Anal Biochem. 2009 Feb 15;385(2):300-8
references.pmid: 19059193
references.citation: Nat Chem Biol. 2006 Feb;2(2):95-102
references.pmid: 16415863
references.citation: Oncogene. 2007 Oct 25;26(49):6989-96
references.pmid: 17486070
references.citation: J Cell Biol. 2004 May 10;165(3):407-19
references.pmid: 15138293
references.citation: Oncogene. 2007 Nov 15;26(52):7313-23
references.pmid: 17533370
references.citation: Adv Cancer Res. 2002;85:51-100
references.pmid: 12374288
references.citation: J Cell Sci. 2003 Jul 1;116(Pt 13):2613-26
references.pmid: 12775773
references.citation: Nat Rev Cancer. 2007 May;7(5):345-56
references.pmid: 17457302
references.citation: Cell. 2003 Mar 21;112(6):845-57
references.pmid: 12654250
references.citation: Oncogene. 2008 Feb 14;27(8):1095-105
references.pmid: 17700528
references.citation: Biochemistry. 1995 Nov 14;34(45):14843-51
references.pmid: 7578094
references.citation: Mol Cell Biol. 2001 Sep;21(18):6233-42
references.pmid: 11509666
references.citation: Cell. 1993 Nov 19;75(4):615-30
references.pmid: 8242738
references.citation: Proc Natl Acad Sci U S A. 1990 Aug;87(15):5802-6
references.pmid: 2198571
references.citation: J Biol Chem. 1985 Jul 5;260(13):8070-7
references.pmid: 2989275
references.citation: Protein Sci. 2005 Dec;14(12):3135-9
references.pmid: 16260764
references.citation: Mol Cell. 2006 Mar 17;21(6):787-98
references.pmid: 16543148
references.citation: Nat Rev Mol Cell Biol. 2004 Jan;5(1):33-44
references.pmid: 14708008