A novel mode of Gleevec binding is revealed by the structure of spleen tyrosine kinase Journal Article
| Authors: | Atwell, S.; Adams, J. M.; Badger, J.; Buchanan, M. D.; Feil, I. K.; Froning, K. J.; Gao, X.; Hendle, J.; Keegan, K.; Leon, B. C.; Müller-Dieckmann, H. J.; Nienaber, V. L.; Noland, B. W.; Post, K.; Rajashankar, K. R.; Ramos, A.; Russell, M.; Burley, S. K.; Buchanan, S. G. |
| Article Title: | A novel mode of Gleevec binding is revealed by the structure of spleen tyrosine kinase |
| Abstract: | Spleen tyrosine kinase (Syk) is a non-receptor tyrosine kinase required for signaling from immunoreceptors in various hematopoietic cells. Phosphorylation of two tyrosine residues in the activation loop of the Syk kinase catalytic domain is necessary for signaling, a phenomenon typical of tyrosine kinase family members. Syk in vitro enzyme activity, however, does not depend on phosphorylation (activation loop tyrosine → phenylalanine mutants retain catalytic activity). We have determined the x-ray structure of the unphosphorylated form of the kinase catalytic domain of Syk. The enzyme adopts a conformation of the activation loop typically seen only in activated, phosphorylated tyrosine kinases, explaining why Syk does not require phosphorylation for activation. We also demonstrate that Gleevec (STI-571, Imatinib) inhibits the isolated kinase domains of both unphosphorylated Syk and phosphorylated Abl with comparable potency. Gleevec binds Syk in a novel, compact cis-conformaiion that differs dramatically from the binding mode observed with unphosphorylated Abl, the more Gleevec-sensitive form of Abl. This finding suggests the existence of two distinct Gleevec binding modes: an extended, trans-conformation characteristic of tight binding to the inactive conformation of a protein kinase and a second compact, cis-conformation characteristic of weaker binding to the active conformation. Finally, the Syk-bound cis-conformation of Gleevec bears a striking resemblance to the rigid structure of the nonspecific, natural product kinase inhibitor staurosporine. |
| Keywords: | signal transduction; protein phosphorylation; mutation; nonhuman; binding affinity; protein conformation; protein domain; animal cell; animals; imatinib; amino acid substitution; protein binding; drug potency; drug structure; enzyme activation; enzyme activity; abelson kinase; pyrimidines; tyrosine; phosphorylation; animalia; immunology; intracellular signaling peptides and proteins; hematopoietic cell; hematopoietic stem cells; ligands; hydrogen bonding; models, molecular; crystallography, x-ray; protein structure, tertiary; binding sites; protein-tyrosine kinases; piperazines; catalysis; enzyme structure; catalytic domain; protein kinase; drug protein binding; enzymes; enzyme precursors; phenylalanine; enzyme conformation; x ray analysis; conformations; cells; x-rays; insects; drug conformation; staurosporine; reaction kinetics; protein kinase syk; structure (composition); humans; priority journal; article; gleevec binding; immunoreceptors; spleen tyrosine kinase (syk) |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 279 |
| Issue: | 53 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2004-12-31 |
| Start Page: | 55827 |
| End Page: | 55832 |
| Language: | English |
| DOI: | 10.1074/jbc.M409792200 |
| PROVIDER: | scopus |
| PUBMED: | 15507431 |
| DOI/URL: | |
| Notes: | J. Biol. Chem. -- Cited By (since 1996):120 -- Export Date: 16 June 2014 -- CODEN: JBCHA -- Source: Scopus |
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