2cww: Difference between revisions

Revision as of 17:53, 21 February 2008

Crystal structure of Thermus thermophilus TTHA1280, a putative SAM-dependent RNA methyltransferase, in complex with S-adenosyl-L-homocysteine

OverviewOverview

The Thermus thermophilus hypothetical protein TTHA1280 belongs to a family of predicted S-adenosyl-L-methionine (AdoMet) dependent RNA methyltransferases (MTases) present in many bacterial and archaeal species. Inspection of amino-acid sequence motifs common to class I Rossmann-fold-like MTases suggested a specific role as an RNA 5-methyluridine MTase. Selenomethionine (SeMet) labelled and native versions of the protein were expressed, purified and crystallized. Two crystal forms of the SeMet-labelled apoprotein were obtained: SeMet-ApoI and SeMet-ApoII. Cocrystallization of the native protein with S-adenosyl-L-homocysteine (AdoHcy) yielded a third crystal form, Native-AdoHcy. The SeMet-ApoI structure was solved by the multiple anomalous dispersion method and refined at 2.55 A resolution. The SeMet-ApoII and Native-AdoHcy structures were solved by molecular replacement and refined at 1.80 and 2.60 A, respectively. TTHA1280 formed a homodimer in the crystals and in solution. Each subunit folds into a three-domain structure composed of a small N-terminal PUA domain, a central alpha/beta-domain and a C-terminal Rossmann-fold-like MTase domain. The three domains form an overall clamp-like shape, with the putative active site facing a deep cleft. The architecture of the active site is consistent with specific recognition of uridine and catalysis of methyl transfer to the 5-carbon position. The cleft is suitable in size and charge distribution for binding single-stranded RNA.

About this StructureAbout this Structure

2CWW is a Single protein structure of sequence from Thermus thermophilus with , and as ligands. Full crystallographic information is available from OCA.

ReferenceReference

Structures of a putative RNA 5-methyluridine methyltransferase, Thermus thermophilus TTHA1280, and its complex with S-adenosyl-L-homocysteine., Pioszak AA, Murayama K, Nakagawa N, Ebihara A, Kuramitsu S, Shirouzu M, Yokoyama S, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Oct 1;61(Pt, 10):867-74. Epub 2005 Sep 30. PMID:16511182

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 ==Overview==
-The Thermus thermophilus hypothetical protein TTHA1280 belongs to a family, of predicted S-adenosyl-L-methionine (AdoMet) dependent RNA, methyltransferases (MTases) present in many bacterial and archaeal, species. Inspection of amino-acid sequence motifs common to class I, Rossmann-fold-like MTases suggested a specific role as an RNA, 5-methyluridine MTase. Selenomethionine (SeMet) labelled and native, versions of the protein were expressed, purified and crystallized. Two, crystal forms of the SeMet-labelled apoprotein were obtained: SeMet-ApoI, and SeMet-ApoII. Cocrystallization of the native protein with, S-adenosyl-L-homocysteine (AdoHcy) yielded a third crystal form, Native-AdoHcy. The SeMet-ApoI structure was solved by the multiple, anomalous dispersion method and refined at 2.55 A resolution. The, SeMet-ApoII and Native-AdoHcy structures were solved by molecular, replacement and refined at 1.80 and 2.60 A, respectively. TTHA1280 formed, a homodimer in the crystals and in solution. Each subunit folds into a, three-domain structure composed of a small N-terminal PUA domain, a, central alpha/beta-domain and a C-terminal Rossmann-fold-like MTase, domain. The three domains form an overall clamp-like shape, with the, putative active site facing a deep cleft. The architecture of the active, site is consistent with specific recognition of uridine and catalysis of, methyl transfer to the 5-carbon position. The cleft is suitable in size, and charge distribution for binding single-stranded RNA.
+The Thermus thermophilus hypothetical protein TTHA1280 belongs to a family of predicted S-adenosyl-L-methionine (AdoMet) dependent RNA methyltransferases (MTases) present in many bacterial and archaeal species. Inspection of amino-acid sequence motifs common to class I Rossmann-fold-like MTases suggested a specific role as an RNA 5-methyluridine MTase. Selenomethionine (SeMet) labelled and native versions of the protein were expressed, purified and crystallized. Two crystal forms of the SeMet-labelled apoprotein were obtained: SeMet-ApoI and SeMet-ApoII. Cocrystallization of the native protein with S-adenosyl-L-homocysteine (AdoHcy) yielded a third crystal form, Native-AdoHcy. The SeMet-ApoI structure was solved by the multiple anomalous dispersion method and refined at 2.55 A resolution. The SeMet-ApoII and Native-AdoHcy structures were solved by molecular replacement and refined at 1.80 and 2.60 A, respectively. TTHA1280 formed a homodimer in the crystals and in solution. Each subunit folds into a three-domain structure composed of a small N-terminal PUA domain, a central alpha/beta-domain and a C-terminal Rossmann-fold-like MTase domain. The three domains form an overall clamp-like shape, with the putative active site facing a deep cleft. The architecture of the active site is consistent with specific recognition of uridine and catalysis of methyl transfer to the 5-carbon position. The cleft is suitable in size and charge distribution for binding single-stranded RNA.
 ==About this Structure==
@@ Line 10: / Line 10: @@
 ==Reference==
-Structures of a putative RNA 5-methyluridine methyltransferase, Thermus thermophilus TTHA1280, and its complex with S-adenosyl-L-homocysteine., Pioszak AA, Murayama K, Nakagawa N, Ebihara A, Kuramitsu S, Shirouzu M, Yokoyama S, Acta Crystallograph Sect F Struct Biol Cryst Commun. 2005 Oct 1;61(Pt, 10):867-74. Epub 2005 Sep 30. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=16511182 16511182]
+Structures of a putative RNA 5-methyluridine methyltransferase, Thermus thermophilus TTHA1280, and its complex with S-adenosyl-L-homocysteine., Pioszak AA, Murayama K, Nakagawa N, Ebihara A, Kuramitsu S, Shirouzu M, Yokoyama S, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Oct 1;61(Pt, 10):867-74. Epub 2005 Sep 30. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=16511182 16511182]
 [[Category: Single protein]]
 [[Category: Thermus thermophilus]]
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 [[Category: Murayama, K.]]
 [[Category: Nakagawa, N.]]
-[[Category: Pioszak, A.A.]]
+[[Category: Pioszak, A A.]]
-[[Category: RSGI, RIKEN.Structural.Genomics/Proteomics.Initiative.]]
+[[Category: RSGI, RIKEN Structural Genomics/Proteomics Initiative.]]
 [[Category: Shirouzu, M.]]
 [[Category: Yokoyama, S.]]
@@ Line 32: / Line 32: @@
 [[Category: structural genomics]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jan 29 18:50:06 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:53:11 2008''