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| A chemical switch for inhibitor-sensitive alleles of any protein kinase |
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| Author(s): Bishop AC, Ubersax JA, Petsch DT, Matheos DP, Gray NS, Blethrow J, Shimizu E, Tsien JZ, Schultz PG, Rose MD, Wood JL, Morgan DO, Shokat KM |
| Source: NATURE Volume: 407 Issue: 6802 Pages: 395-401 Published: SEP 21 2000 |
| Times Cited: 301 References: 30 |
| Abstract: Protein kinases have proved to be largely resistant to the design of highly specific inhibitors, even with the aid of combinatorial chemistry(1). The lack of these reagents has complicated efforts to assign specific signalling roles to individual kinases. Here we describe a chemical genetic strategy for sensitizing protein kinases to cell-permeable molecules that do not inhibit wildtype kinases(2). From two inhibitor scaffolds, we have identified potent and selective inhibitors for sensitized kinases from five distinct subfamilies. Tyrosine and serine/threonine kinases are equally amenable to this approach. We have analysed a budding yeast strain carrying an inhibitor-sensitive form of the cyclin-dependent kinase Cdc28 (CDK1) in place of the wild-type protein. Specific inhibition of Cdc28 in vivo caused a pre-mitotic cell-cycle arrest that is distinct from the G1 arrest typically observed in temperature-sensitive cdc28 mutants(3). The mutation that confers inhibitor-sensitivity is easily identifiable from primary sequence alignments. Thus, this approach can be used to systematically generate conditional alleles of protein kinases, allowing for rapid functional characterization of members of this important gene family. |
| Document Type: Article |
| Language: English |
| Reprint Address: Shokat, KM (reprint author), Univ Calif San Francisco, Dept Mol & Cellular Pharmacol, San Francisco, CA 94143 USA |
Addresses:
1. Univ Calif San Francisco, Dept Physiol, San Francisco, CA 94143 USA
2. Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94143 USA
3. Yale Univ, Dept Chem, New Haven, CT 06520 USA
4. Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA
5. Novartis Res Fdn, Genom Inst, San Diego, CA 92121 USA
6. Princeton Univ, Dept Chem, Princeton, NJ 08544 USA |
| Publisher: MACMILLAN PUBLISHERS LTD, PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND |
| Subject Category: Multidisciplinary Sciences |
| IDS Number: 355NA |
| ISSN: 0028-0836 |
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