2ivu
From Proteopedia
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==Overview== | ==Overview== | ||
| - | The RET proto-oncogene encodes a receptor tyrosine kinase for the glial, cell line-derived neurotrophic factor family of ligands. Loss-of-function, mutations in RET are implicated in Hirschsprung disease, whereas, activating mutations in RET are found in human cancers, including familial, medullar thyroid carcinoma and multiple endocrine neoplasias 2A and 2B. We, report here the biochemical characterization of the human RET tyrosine, kinase domain and the structure determination of the non-phosphorylated, and phosphorylated forms. Both structures adopt the same active kinase, conformation competent to bind ATP and substrate and have a pre-organized, activation loop conformation that is independent of phosphorylation, status. In agreement with the structural data, enzyme kinetic data show, ... | + | The RET proto-oncogene encodes a receptor tyrosine kinase for the glial, cell line-derived neurotrophic factor family of ligands. Loss-of-function, mutations in RET are implicated in Hirschsprung disease, whereas, activating mutations in RET are found in human cancers, including familial, medullar thyroid carcinoma and multiple endocrine neoplasias 2A and 2B. We, report here the biochemical characterization of the human RET tyrosine, kinase domain and the structure determination of the non-phosphorylated, and phosphorylated forms. Both structures adopt the same active kinase, conformation competent to bind ATP and substrate and have a pre-organized, activation loop conformation that is independent of phosphorylation, status. In agreement with the structural data, enzyme kinetic data show, that autophosphorylation produces only a modest increase in activity., Longer forms of RET containing the juxtamembrane domain and C-terminal, tail exhibited similar kinetic behavior, implying that there is no, cis-inhibitory mechanism within the RET intracellular domain. Our results, suggest the existence of alternative inhibitory mechanisms, possibly in, trans, for the autoregulation of RET kinase activity. We also present the, structures of the RET tyrosine kinase domain bound to two inhibitors, the, pyrazolopyrimidine PP1 and the clinically relevant 4-anilinoquinazoline, ZD6474. These structures explain why certain multiple endocrine neoplasia, 2-associated RET mutants found in patients are resistant to inhibition and, form the basis for design of more effective inhibitors. |
==About this Structure== | ==About this Structure== | ||
| - | 2IVU is a | + | 2IVU is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with ZD6 and FMT as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Receptor_protein-tyrosine_kinase Receptor protein-tyrosine kinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.10.1 2.7.10.1] Structure known Active Site: AC1. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2IVU OCA]. |
==Reference== | ==Reference== | ||
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[[Category: tyrosine-protein kinase]] | [[Category: tyrosine-protein kinase]] | ||
| - | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 13:47:14 2007'' |
Revision as of 11:41, 5 November 2007
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CRYSTAL STRUCTURE OF PHOSPHORYLATED RET TYROSINE KINASE DOMAIN COMPLEXED WITH THE INHIBITOR ZD6474
Overview
The RET proto-oncogene encodes a receptor tyrosine kinase for the glial, cell line-derived neurotrophic factor family of ligands. Loss-of-function, mutations in RET are implicated in Hirschsprung disease, whereas, activating mutations in RET are found in human cancers, including familial, medullar thyroid carcinoma and multiple endocrine neoplasias 2A and 2B. We, report here the biochemical characterization of the human RET tyrosine, kinase domain and the structure determination of the non-phosphorylated, and phosphorylated forms. Both structures adopt the same active kinase, conformation competent to bind ATP and substrate and have a pre-organized, activation loop conformation that is independent of phosphorylation, status. In agreement with the structural data, enzyme kinetic data show, that autophosphorylation produces only a modest increase in activity., Longer forms of RET containing the juxtamembrane domain and C-terminal, tail exhibited similar kinetic behavior, implying that there is no, cis-inhibitory mechanism within the RET intracellular domain. Our results, suggest the existence of alternative inhibitory mechanisms, possibly in, trans, for the autoregulation of RET kinase activity. We also present the, structures of the RET tyrosine kinase domain bound to two inhibitors, the, pyrazolopyrimidine PP1 and the clinically relevant 4-anilinoquinazoline, ZD6474. These structures explain why certain multiple endocrine neoplasia, 2-associated RET mutants found in patients are resistant to inhibition and, form the basis for design of more effective inhibitors.
About this Structure
2IVU is a Single protein structure of sequence from Homo sapiens with ZD6 and FMT as ligands. Active as Receptor protein-tyrosine kinase, with EC number 2.7.10.1 Structure known Active Site: AC1. Full crystallographic information is available from OCA.
Reference
Structure and chemical inhibition of the RET tyrosine kinase domain., Knowles PP, Murray-Rust J, Kjaer S, Scott RP, Hanrahan S, Santoro M, Ibanez CF, McDonald NQ, J Biol Chem. 2006 Nov 3;281(44):33577-87. Epub 2006 Aug 23. PMID:16928683
Page seeded by OCA on Mon Nov 5 13:47:14 2007
Categories: Homo sapiens | Receptor protein-tyrosine kinase | Single protein | Knowles, P.P. | Mcdonald, N.Q. | Murray-Rust, J. | FMT | ZD6 | Atp-binding | Chromosomal translocation | Disease mutation | Gdnf receptor | Hirschsprung disease | Kinase | Membrane | Nucleotide-binding | Phosphorylation | Phosphotransferase | Polymorphism | Proto-oncogene | Ret | Transferase | Transmembrane | Tyrosine kinase | Tyrosine-protein kinase
