4LX3

Conserved Residues that Modulate Protein trans-Splicing of Npu DnaE Split Intein

4LX3 の概要

• エントリーDOI: 10.2210/pdb4lx3/pdb
• 分子名称: DNA polymerase III, alpha subunit, Nucleic acid binding, OB-fold, tRNA/helicase-type (3 entities in total)
• 機能のキーワード: trans-structure, naturally occurring, transferase
• 由来する生物種: Nostoc punctiforme; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 17016.61

構造登録者

Wu, Q.,Gao, Z.,Wei, Y.,Ma, G.,Zheng, Y.,Dong, Y.,Liu, Y. (登録日: 2013-07-29, 公開日: 2014-06-25, 最終更新日: 2024-11-13)

主引用文献

Wu, Q.,Gao, Z.,Wei, Y.,Ma, G.,Zheng, Y.,Dong, Y.,Liu, Y.
Conserved residues that modulate protein trans-splicing of Npu DnaE split intein.
Biochem.J., 461:247-255, 2014

PubMed Abstract: The first crystal trans-structure of a naturally occurring split intein has been determined for the Npu (Nostoc punctiforme PCC73102) DnaE split intein. Guided by this structure, the residues NArg50 and CSer35, well conserved in DnaE split inteins, are identified to be critical in the trans-splicing of Npu DnaE split intein. An in vitro splicing assay demonstrates that NArg50 and CSer35 play synergistic roles in modulating its intein activity. The C-terminal CAsn36 exhibits two orientations of its side chain and interacts with both NArg50 and CSer35 through hydrogen bonding. These interactions likely facilitate the cyclization of asparagine in the course of protein splicing. The mutation of either residue reduces intein activity, and correlates with the low activity of the Ssp (Cyanobacterium synechocystis sp. strain PCC6803) DnaE split intein. On the other hand, NArg50 also forms a hydrogen bond with the highly conserved F-block CAsp17, thus influencing the N-S acyl shift during N-terminal cleavage. Sequence alignments show that residues NArg50 and CSer35 are rather conserved in those split inteins that lack a penultimate histidine residue. The conserved non-catalytic residues of split inteins modulate the efficiency of protein trans-splicing by hydrogen-bond interactions with the catalytic residues at the splice junction.

PubMed: 24758175
DOI: 10.1042/BJ20140287

実験手法

X-RAY DIFFRACTION (1.5 Å)