2FYH

Solution structure of the 2'-5' RNA ligase-like protein from Pyrococcus furiosus

2FYH の概要

• エントリーDOI: 10.2210/pdb2fyh/pdb
• 分子名称: putative integral membrane transport protein (1 entity in total)
• 機能のキーワード: 2'-5' rna ligase-like protein, hxtx motif, pyrococcus furiosus, structural genomics, nppsfa, national project on protein structural and functional analyses, riken structural genomics/proteomics initiative, rsgi, ligase
• 由来する生物種: Pyrococcus furiosus
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 21860.40

構造登録者

Okada, K.,Matsuda, T.,Sakamoto, T.,Muto, Y.,Yokoyama, S.,Kanai, A.,Kawai, G.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (登録日: 2006-02-08, 公開日: 2007-02-20, 最終更新日: 2024-05-01)

主引用文献

Kanai, A.,Sato, A.,Fukuda, Y.,Okada, K.,Matsuda, T.,Sakamoto, T.,Muto, Y.,Yokoyama, S.,Kawai, G.,Tomita, M.
Characterization of a heat-stable enzyme possessing GTP-dependent RNA ligase activity from a hyperthermophilic archaeon, Pyrococcus furiosus
Rna, 15:420-431, 2009

PubMed Abstract: Using an expression protein library of a hyperthermophilic archaeon, Pyrococcus furiosus, we identified a gene (PF0027) that encodes a protein with heat-stable cyclic nucleotide phosphodiesterase (CPDase) activity. The PF0027 gene encoded a 21-kDa protein and an amino acid sequence that showed approximately 27% identity to that of the 2'-5' tRNA ligase protein, ligT (20 kDa), from Escherichia coli. We found that the purified PF0027 protein possessed GTP-dependent RNA ligase activity and that synthetic tRNA halves bearing 2',3'-cyclic phosphate and 5'-OH termini were substrates for the ligation reaction in vitro. GTP hydrolysis was not required for the reaction, and GTPgammaS enhanced the tRNA ligation activity of PF0027 protein, suggesting that the ligation step is regulated by a novel mechanism. In comparison to the strong CPDase activity of the PF0027 protein, the RNA ligase activity itself was quite weak, and the ligation product was unstable during in vitro reaction. Finally, we used NMR to determine the solution structure of the PF0027 protein and discuss the implications of our results in understanding the role of the PF0027 protein.

PubMed: 19155324
DOI: 10.1261/rna.1122109

実験手法

SOLUTION NMR