2i3b
From Proteopedia
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==Overview== | ==Overview== | ||
| - | A screen of the human cancer genome anatomy project (CGAP) database was | + | A screen of the human cancer genome anatomy project (CGAP) database was performed to search for new proteins involved in tumorigenesis. The resulting hits were further screened for recombinant expression, solubility and protein aggregation, which led to the identification of the previously unknown human cancer-related (HCR) protein encoded by the mRNA NM_032324 as a target for structure determination by NMR. The three-dimensional structure of the protein in its complex with ATPgammaS forms a three-layered alpha/beta sandwich, with a central nine-stranded beta-sheet surrounded by five alpha-helices. Sequence and three-dimensional structure comparisons with AAA+ ATPases revealed the presence of Walker A (GPPGVGKT) and Walker B (VCVIDEIG) motifs. Using 1D (31)P-NMR spectroscopy and a coupled enzymatic assay for the determination of inorganic phosphate, we showed that the purified recombinant protein is active as a non-specific nucleoside triphosphatase, with k(cat)=7.6x10(-3) s(-1). The structural basis for the enzymatic activity of HCR-NTPase was further characterized by site-directed mutagenesis of the Walker B motif, which further contributes to making the HCR-NTPase an attractive new target for further biochemical characterization in the context of its presumed role in human tumorigenesis. |
==About this Structure== | ==About this Structure== | ||
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==Reference== | ==Reference== | ||
| - | NMR | + | NMR structure and functional characterization of a human cancer-related nucleoside triphosphatase., Placzek WJ, Almeida MS, Wuthrich K, J Mol Biol. 2007 Mar 30;367(3):788-801. Epub 2007 Jan 9. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17291528 17291528] |
[[Category: Homo sapiens]] | [[Category: Homo sapiens]] | ||
[[Category: Nucleoside-triphosphatase]] | [[Category: Nucleoside-triphosphatase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Almeida, M | + | [[Category: Almeida, M S.]] |
| - | [[Category: Placzek, W | + | [[Category: Placzek, W J.]] |
[[Category: Wuthrich, K.]] | [[Category: Wuthrich, K.]] | ||
[[Category: aaa]] | [[Category: aaa]] | ||
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[[Category: rossmann]] | [[Category: rossmann]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:48:36 2008'' |
Revision as of 15:48, 21 February 2008
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Solution Structure of a Human Cancer-Related Nucleoside Triphosphatase
Overview
A screen of the human cancer genome anatomy project (CGAP) database was performed to search for new proteins involved in tumorigenesis. The resulting hits were further screened for recombinant expression, solubility and protein aggregation, which led to the identification of the previously unknown human cancer-related (HCR) protein encoded by the mRNA NM_032324 as a target for structure determination by NMR. The three-dimensional structure of the protein in its complex with ATPgammaS forms a three-layered alpha/beta sandwich, with a central nine-stranded beta-sheet surrounded by five alpha-helices. Sequence and three-dimensional structure comparisons with AAA+ ATPases revealed the presence of Walker A (GPPGVGKT) and Walker B (VCVIDEIG) motifs. Using 1D (31)P-NMR spectroscopy and a coupled enzymatic assay for the determination of inorganic phosphate, we showed that the purified recombinant protein is active as a non-specific nucleoside triphosphatase, with k(cat)=7.6x10(-3) s(-1). The structural basis for the enzymatic activity of HCR-NTPase was further characterized by site-directed mutagenesis of the Walker B motif, which further contributes to making the HCR-NTPase an attractive new target for further biochemical characterization in the context of its presumed role in human tumorigenesis.
About this Structure
2I3B is a Single protein structure of sequence from Homo sapiens. Active as Nucleoside-triphosphatase, with EC number 3.6.1.15 Full crystallographic information is available from OCA.
Reference
NMR structure and functional characterization of a human cancer-related nucleoside triphosphatase., Placzek WJ, Almeida MS, Wuthrich K, J Mol Biol. 2007 Mar 30;367(3):788-801. Epub 2007 Jan 9. PMID:17291528
Page seeded by OCA on Thu Feb 21 17:48:36 2008
