1q2s
From Proteopedia
(New page: 200px<br /><applet load="1q2s" size="450" color="white" frame="true" align="right" spinBox="true" caption="1q2s, resolution 3.2Å" /> '''Chemical trapping and...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:1q2s.gif|left|200px]]<br /><applet load="1q2s" size=" | + | [[Image:1q2s.gif|left|200px]]<br /><applet load="1q2s" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1q2s, resolution 3.2Å" /> | caption="1q2s, resolution 3.2Å" /> | ||
'''Chemical trapping and crystal structure of a catalytic tRNA guanine transglycosylase covalent intermediate'''<br /> | '''Chemical trapping and crystal structure of a catalytic tRNA guanine transglycosylase covalent intermediate'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Prokaryotic tRNA guanine transglycosylase (TGT) catalyzes replacement of | + | Prokaryotic tRNA guanine transglycosylase (TGT) catalyzes replacement of guanine (G) by 7-aminomethyl-7-deazaguanine (PreQ1) at the wobble position of four specific tRNAs. Addition of 9-deazaguanine (9dzG) to a reaction mixture of Zymomonas mobilis TGT and an RNA substrate allowed us to trap, purify and crystallize a chemically competent covalent intermediate of the TGT-catalyzed reaction. The crystal structure of the TGT-RNA-9dzG ternary complex at a resolution of 2.9 A reveals, unexpectedly, that RNA is tethered to TGT through the side chain of Asp280. Thus, Asp280, instead of the previously proposed Asp102, acts as the nucleophile for the reaction. The RNA substrate adopts an unusual conformation, with four out of seven nucleotides in the loop region flipped out. Interactions between TGT and RNA revealed by the structure provide the molecular basis of the RNA substrate requirements by TGT. Furthermore, reaction of PreQ1 with the crystallized covalent intermediate provides insight into the necessary structural changes required for the TGT-catalyzed reaction to occur. |
==About this Structure== | ==About this Structure== | ||
| - | 1Q2S is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Zymomonas_mobilis Zymomonas mobilis] with ZN and 9DG as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Queuine_tRNA-ribosyltransferase Queuine tRNA-ribosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.2.29 2.4.2.29] Full crystallographic information is available from [http:// | + | 1Q2S is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Zymomonas_mobilis Zymomonas mobilis] with <scene name='pdbligand=ZN:'>ZN</scene> and <scene name='pdbligand=9DG:'>9DG</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Queuine_tRNA-ribosyltransferase Queuine tRNA-ribosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.2.29 2.4.2.29] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1Q2S OCA]. |
==Reference== | ==Reference== | ||
| Line 14: | Line 14: | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Zymomonas mobilis]] | [[Category: Zymomonas mobilis]] | ||
| - | [[Category: Huang, R | + | [[Category: Huang, R H.]] |
[[Category: Liu, X.]] | [[Category: Liu, X.]] | ||
[[Category: Xie, W.]] | [[Category: Xie, W.]] | ||
| Line 23: | Line 23: | ||
[[Category: tim barrel]] | [[Category: tim barrel]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:35:19 2008'' |
Revision as of 12:35, 21 February 2008
|
Chemical trapping and crystal structure of a catalytic tRNA guanine transglycosylase covalent intermediate
Overview
Prokaryotic tRNA guanine transglycosylase (TGT) catalyzes replacement of guanine (G) by 7-aminomethyl-7-deazaguanine (PreQ1) at the wobble position of four specific tRNAs. Addition of 9-deazaguanine (9dzG) to a reaction mixture of Zymomonas mobilis TGT and an RNA substrate allowed us to trap, purify and crystallize a chemically competent covalent intermediate of the TGT-catalyzed reaction. The crystal structure of the TGT-RNA-9dzG ternary complex at a resolution of 2.9 A reveals, unexpectedly, that RNA is tethered to TGT through the side chain of Asp280. Thus, Asp280, instead of the previously proposed Asp102, acts as the nucleophile for the reaction. The RNA substrate adopts an unusual conformation, with four out of seven nucleotides in the loop region flipped out. Interactions between TGT and RNA revealed by the structure provide the molecular basis of the RNA substrate requirements by TGT. Furthermore, reaction of PreQ1 with the crystallized covalent intermediate provides insight into the necessary structural changes required for the TGT-catalyzed reaction to occur.
About this Structure
1Q2S is a Single protein structure of sequence from Zymomonas mobilis with and as ligands. Active as Queuine tRNA-ribosyltransferase, with EC number 2.4.2.29 Full crystallographic information is available from OCA.
Reference
Chemical trapping and crystal structure of a catalytic tRNA guanine transglycosylase covalent intermediate., Xie W, Liu X, Huang RH, Nat Struct Biol. 2003 Oct;10(10):781-8. Epub 2003 Aug 31. PMID:12949492
Page seeded by OCA on Thu Feb 21 14:35:19 2008
