1nht
From Proteopedia
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==Overview== | ==Overview== | ||
| - | Crystal structures of adenylosuccinate synthetase from Escherichia coli, complexed with Mg2+, 6-thiophosphoryl-IMP, GDP, and hadacidin at 298 and, 100 K have been refined to R-factors of 0.171 and 0.206 against data to, 2.8 and 2.5 A resolution, respectively. Interactions of GDP, Mg2+ and, hadacidin are similar to those observed for the same ligands in the, complex of IMP, GDP, NO3-, Mg2+ and hadacidin (Poland, B. W., Fromm, H. J., & Honzatko, R. B. (1996). J. Mol. Biol. 264, 1013-1027). Although crystals, were grown from solutions containing 6-mercapto-IMP and GTP, the electron, density at the active site is consistent with 6-thiophosphoryl-IMP and, GDP. Asp-13 and Gln-224 probably work in concert to stabilize the, 6-thioanion of 6-mercapto-IMP, which in turn is the nucleophile in ... | + | Crystal structures of adenylosuccinate synthetase from Escherichia coli, complexed with Mg2+, 6-thiophosphoryl-IMP, GDP, and hadacidin at 298 and, 100 K have been refined to R-factors of 0.171 and 0.206 against data to, 2.8 and 2.5 A resolution, respectively. Interactions of GDP, Mg2+ and, hadacidin are similar to those observed for the same ligands in the, complex of IMP, GDP, NO3-, Mg2+ and hadacidin (Poland, B. W., Fromm, H. J., & Honzatko, R. B. (1996). J. Mol. Biol. 264, 1013-1027). Although crystals, were grown from solutions containing 6-mercapto-IMP and GTP, the electron, density at the active site is consistent with 6-thiophosphoryl-IMP and, GDP. Asp-13 and Gln-224 probably work in concert to stabilize the, 6-thioanion of 6-mercapto-IMP, which in turn is the nucleophile in the, displacement of GDP from the gamma-phosphate of GTP. Once formed, 6-thiophosphoryl-IMP is stable in the active site of the enzyme under the, conditions of the structural investigation. The direct observation of, 6-thiophosphoryl-IMP in the active site is consistent with the putative, generation of 6-phosphoryl-IMP along the reaction pathway of the, synthetase. |
==About this Structure== | ==About this Structure== | ||
| - | 1NHT is a | + | 1NHT is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with MG, GDP, HAD and PGS as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Adenylosuccinate_synthase Adenylosuccinate synthase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=6.3.4.4 6.3.4.4] Structure known Active Sites: ASP, GNS and IMP. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1NHT OCA]. |
==Reference== | ==Reference== | ||
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[[Category: x-ray crystallography]] | [[Category: x-ray crystallography]] | ||
| - | ''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 13:58:55 2007'' |
Revision as of 11:53, 5 November 2007
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ENTRAPMENT OF 6-THIOPHOSPHORYL-IMP IN THE ACTIVE SITE OF CRYSTALLINE ADENYLOSUCCINATE SYNTHETASE FROM ESCHERICHIA COLI DATA COLLECTED AT 100K
Overview
Crystal structures of adenylosuccinate synthetase from Escherichia coli, complexed with Mg2+, 6-thiophosphoryl-IMP, GDP, and hadacidin at 298 and, 100 K have been refined to R-factors of 0.171 and 0.206 against data to, 2.8 and 2.5 A resolution, respectively. Interactions of GDP, Mg2+ and, hadacidin are similar to those observed for the same ligands in the, complex of IMP, GDP, NO3-, Mg2+ and hadacidin (Poland, B. W., Fromm, H. J., & Honzatko, R. B. (1996). J. Mol. Biol. 264, 1013-1027). Although crystals, were grown from solutions containing 6-mercapto-IMP and GTP, the electron, density at the active site is consistent with 6-thiophosphoryl-IMP and, GDP. Asp-13 and Gln-224 probably work in concert to stabilize the, 6-thioanion of 6-mercapto-IMP, which in turn is the nucleophile in the, displacement of GDP from the gamma-phosphate of GTP. Once formed, 6-thiophosphoryl-IMP is stable in the active site of the enzyme under the, conditions of the structural investigation. The direct observation of, 6-thiophosphoryl-IMP in the active site is consistent with the putative, generation of 6-phosphoryl-IMP along the reaction pathway of the, synthetase.
About this Structure
1NHT is a Single protein structure of sequence from Escherichia coli with MG, GDP, HAD and PGS as ligands. Active as Adenylosuccinate synthase, with EC number 6.3.4.4 Structure known Active Sites: ASP, GNS and IMP. Full crystallographic information is available from OCA.
Reference
Entrapment of 6-thiophosphoryl-IMP in the active site of crystalline adenylosuccinate synthetase from Escherichia coli., Poland BW, Bruns C, Fromm HJ, Honzatko RB, J Biol Chem. 1997 Jun 13;272(24):15200-5. PMID:9182542
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