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New page: left|200px<br /><applet load="2aae" size="450" color="white" frame="true" align="right" spinBox="true" caption="2aae, resolution 1.8Å" /> '''THE ROLE OF HISTIDINE...
 
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[[Image:2aae.jpg|left|200px]]<br /><applet load="2aae" size="450" color="white" frame="true" align="right" spinBox="true"  
[[Image:2aae.jpg|left|200px]]<br /><applet load="2aae" size="350" color="white" frame="true" align="right" spinBox="true"  
caption="2aae, resolution 1.8&Aring;" />
caption="2aae, resolution 1.8&Aring;" />
'''THE ROLE OF HISTIDINE-40 IN RIBONUCLEASE T1 CATALYSIS: THREE-DIMENSIONAL STRUCTURES OF THE PARTIALLY ACTIVE HIS40LYS MUTANT'''<br />
'''THE ROLE OF HISTIDINE-40 IN RIBONUCLEASE T1 CATALYSIS: THREE-DIMENSIONAL STRUCTURES OF THE PARTIALLY ACTIVE HIS40LYS MUTANT'''<br />


==Overview==
==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., &amp; 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., &amp; Saenger, W. (1988) J. Biol. Chem. 263, 15358-15368] and vanadate [Kostrewa, D., Hui-Woog Choe, Heinemann, U., &amp;, 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)
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., &amp; 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., &amp; Saenger, W. (1988) J. Biol. Chem. 263, 15358-15368] and vanadate [Kostrewa, D., Hui-Woog Choe, Heinemann, U., &amp; 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==
2AAE 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 and PO4 as [http://en.wikipedia.org/wiki/ligands ligands]. 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] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2AAE OCA].  
2AAE 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 <scene name='pdbligand=CA:'>CA</scene> and <scene name='pdbligand=PO4:'>PO4</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. 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] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2AAE OCA].  


==Reference==
==Reference==
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[[Category: Ribonuclease T(1)]]
[[Category: Ribonuclease T(1)]]
[[Category: Single protein]]
[[Category: Single protein]]
[[Category: Choe, C.W.]]
[[Category: Choe, C W.]]
[[Category: Heinemann, U.]]
[[Category: Heinemann, U.]]
[[Category: Saenger, W.]]
[[Category: Saenger, W.]]
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[[Category: hydrolase(endoribonuclease)]]
[[Category: hydrolase(endoribonuclease)]]


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