5FFJ

Structure of a nuclease-deletion mutant of the Type ISP restriction-modification enzyme LlaGI in complex with a DNA substrate mimic

5FFJ の概要

• エントリーDOI: 10.2210/pdb5ffj/pdb
• 関連するPDBエントリー: 4XQK
• 分子名称: Endonuclease and methylase LlaGI, DNA (5'-D(P*TP*CP*CP*TP*CP*CP*AP*TP*CP*CP*AP*GP*TP*CP*TP*AP*TP*TP*AP*GP*CP*T)-3'), DNA (5'-D(P*TP*AP*GP*CP*TP*AP*AP*TP*AP*GP*AP*CP*TP*GP*GP*AP*TP*GP*GP*AP*GP*G)-3'), ... (4 entities in total)
• 機能のキーワード: helicase-like atpase, methyltransferase, dna-binding protein, restriction-modification enzyme, dna binding protein-dna complex, dna binding protein/dna
• 由来する生物種: Lactococcus lactis; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 349661.66

構造登録者

Saikrishnan, K.,Kulkarni, M.,Nirwan, N. (登録日: 2015-12-18, 公開日: 2016-03-30, 最終更新日: 2024-03-20)

主引用文献

Kulkarni, M.,Nirwan, N.,van Aelst, K.,Szczelkun, M.D.,Saikrishnan, K.
Structural insights into DNA sequence recognition by Type ISP restriction-modification enzymes
Nucleic Acids Res., 44:4396-4408, 2016

PubMed Abstract: Engineering restriction enzymes with new sequence specificity has been an unaccomplished challenge, presumably because of the complexity of target recognition. Here we report detailed analyses of target recognition by Type ISP restriction-modification enzymes. We determined the structure of the Type ISP enzyme LlaGI bound to its target and compared it with the previously reported structure of a close homologue that binds to a distinct target, LlaBIII. The comparison revealed that, although the two enzymes use almost a similar set of structural elements for target recognition, the residues that read the bases vary. Change in specificity resulted not only from appropriate substitution of amino acids that contacted the bases but also from new contacts made by positionally distinct residues directly or through a water bridge. Sequence analyses of 552 Type ISP enzymes showed that the structural elements involved in target recognition of LlaGI and LlaBIII were structurally well-conserved but sequentially less-conserved. In addition, the residue positions within these structural elements were under strong evolutionary constraint, highlighting the functional importance of these regions. The comparative study helped decipher a partial consensus code for target recognition by Type ISP enzymes.

PubMed: 26975655
DOI: 10.1093/nar/gkw154

実験手法

X-RAY DIFFRACTION (2.84 Å)