4bir

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==Overview==
==Overview==
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Histidine-40 is known to participate in phosphodiester transesterification, catalyzed by the enzyme ribonuclease T1. A mutant enzyme with a lysine, replacing the histidine-40 (His40Lys RNase T1) retains considerable, catalytic activity [Steyaert, J., Hallenga, K., Wyns, L., & Stanssens, P., (1990) Biochemistry 29, 9064-9072]. We report on the crystal structures of, His40Lys RNase T1 containing a phosphate anion and a guanosine, 2'-phosphate inhibitor in the active site, respectively. Similar to, previously described structures, the phosphate-containing crystals are of, space group P2(1)2(1)2(1), with one molecule per asymmetric unit (a =, 48.27 A, b = 46.50 A, c = 41.14 A). The complex with 2'-GMP crystallized, in the lower symmetry space group P2(1), with two molecules per ... [[http://ispc.weizmann.ac.il/pmbin/getpm?1445870 (full description)]]
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Histidine-40 is known to participate in phosphodiester transesterification, catalyzed by the enzyme ribonuclease T1. A mutant enzyme with a lysine, replacing the histidine-40 (His40Lys RNase T1) retains considerable, catalytic activity [Steyaert, J., Hallenga, K., Wyns, L., & Stanssens, P., (1990) Biochemistry 29, 9064-9072]. We report on the crystal structures of, His40Lys RNase T1 containing a phosphate anion and a guanosine, 2'-phosphate inhibitor in the active site, respectively. Similar to, previously described structures, the phosphate-containing crystals are of, space group P2(1)2(1)2(1), with one molecule per asymmetric unit (a =, 48.27 A, b = 46.50 A, c = 41.14 A). The complex with 2'-GMP crystallized, in the lower symmetry space group P2(1), with two molecules per asymmetric, unit (a = 49.20 A, b = 48.19 A, c = 40.16 A, beta = 90.26). The crystal, structures have been solved at 1.8- and 2.0-A resolution yielding R values, of 14.5% and 16.0%, respectively. Comparison of these His40Lys structures, with the corresponding wild-type structures, containing 2'-GMP [Arni, R., Heinemann, U., Tokuoka, R., & Saenger, W. (1988) J. Biol. Chem. 263, 15358-15368] and vanadate [Kostrewa, D., Hui-Woog Choe, Heinemann, U., &, Saenger, W. (1989) Biochemistry 28, 7692-7600] in the active site, respectively, leads to the following conclusions. First, the His40Lys, mutation causes no significant changes in the overall structure of RNase, T1; second, the Lys40 side chains in the mutant structures occupy roughly, the same space as His40 in the corresponding wild-type RNase T1, structures.(ABSTRACT TRUNCATED AT 250 WORDS)
==About this Structure==
==About this Structure==
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4BIR is a [[http://en.wikipedia.org/wiki/Single_protein Single protein]] structure of sequence from [[http://en.wikipedia.org/wiki/Aspergillus_oryzae Aspergillus oryzae]] with CA as [[http://en.wikipedia.org/wiki/ligand ligand]]. Active as [[http://en.wikipedia.org/wiki/Ribonuclease_T(1) Ribonuclease T(1)]], with EC number [[http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.27.3 3.1.27.3]]. Structure known Active Site: CAT. Full crystallographic information is available from [[http://ispc.weizmann.ac.il/oca-bin/ocashort?id=4BIR OCA]].
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4BIR is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aspergillus_oryzae Aspergillus oryzae] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Ribonuclease_T(1) Ribonuclease T(1)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.27.3 3.1.27.3] Structure known Active Site: CAT. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=4BIR OCA].
==Reference==
==Reference==
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[[Category: ribonuclease]]
[[Category: ribonuclease]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Oct 30 17:47:22 2007''
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 5 15:21:25 2007''

Revision as of 13:16, 5 November 2007


4bir, resolution 1.70Å

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RIBONUCLEASE T1: FREE HIS92GLN MUTANT

Overview

Histidine-40 is known to participate in phosphodiester transesterification, catalyzed by the enzyme ribonuclease T1. A mutant enzyme with a lysine, replacing the histidine-40 (His40Lys RNase T1) retains considerable, catalytic activity [Steyaert, J., Hallenga, K., Wyns, L., & Stanssens, P., (1990) Biochemistry 29, 9064-9072]. We report on the crystal structures of, His40Lys RNase T1 containing a phosphate anion and a guanosine, 2'-phosphate inhibitor in the active site, respectively. Similar to, previously described structures, the phosphate-containing crystals are of, space group P2(1)2(1)2(1), with one molecule per asymmetric unit (a =, 48.27 A, b = 46.50 A, c = 41.14 A). The complex with 2'-GMP crystallized, in the lower symmetry space group P2(1), with two molecules per asymmetric, unit (a = 49.20 A, b = 48.19 A, c = 40.16 A, beta = 90.26). The crystal, structures have been solved at 1.8- and 2.0-A resolution yielding R values, of 14.5% and 16.0%, respectively. Comparison of these His40Lys structures, with the corresponding wild-type structures, containing 2'-GMP [Arni, R., Heinemann, U., Tokuoka, R., & Saenger, W. (1988) J. Biol. Chem. 263, 15358-15368] and vanadate [Kostrewa, D., Hui-Woog Choe, Heinemann, U., &, Saenger, W. (1989) Biochemistry 28, 7692-7600] in the active site, respectively, leads to the following conclusions. First, the His40Lys, mutation causes no significant changes in the overall structure of RNase, T1; second, the Lys40 side chains in the mutant structures occupy roughly, the same space as His40 in the corresponding wild-type RNase T1, structures.(ABSTRACT TRUNCATED AT 250 WORDS)

About this Structure

4BIR is a Single protein structure of sequence from Aspergillus oryzae with CA as ligand. Active as Ribonuclease T(1), with EC number 3.1.27.3 Structure known Active Site: CAT. Full crystallographic information is available from OCA.

Reference

Role of histidine-40 in ribonuclease T1 catalysis: three-dimensionalstructures of the partially active His40Lys mutant., Zegers I, Verhelst P, Choe HW, Steyaert J, Heinemann U, Saenger W, Wyns L, Biochemistry. 1992 Nov 24;31(46):11317-25. PMID:1445870

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