5HR4

Structure of Type IIL restriction-modification enzyme MmeI in complex with DNA has implications for engineering of new specificities

5HR4 の概要

• エントリーDOI: 10.2210/pdb5hr4/pdb
• 分子名称: MmeI, DNA (5'-D(P*TP*AP*TP*CP*CP*GP*AP*CP*AP*TP*AP*AP*C)-3'), DNA (5'-D(P*GP*TP*TP*AP*TP*GP*TP*CP*GP*GP*AP*TP*A)-3'), ... (6 entities in total)
• 機能のキーワード: dna-protein complex, restriction-modification enzyme, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Methylophilus methylotrophus; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 227254.67

構造登録者

Callahan, S.J.,Luyten, Y.A.,Gupta, Y.K.,Wilson, G.G.,Roberts, R.J.,Morgan, R.D.,Aggarwal, A.K. (登録日: 2016-01-22, 公開日: 2016-04-27, 最終更新日: 2024-03-06)

主引用文献

Callahan, S.J.,Luyten, Y.A.,Gupta, Y.K.,Wilson, G.G.,Roberts, R.J.,Morgan, R.D.,Aggarwal, A.K.
Structure of Type IIL Restriction-Modification Enzyme MmeI in Complex with DNA Has Implications for Engineering New Specificities.
Plos Biol., 14:e1002442-e1002442, 2016

PubMed Abstract: The creation of restriction enzymes with programmable DNA-binding and -cleavage specificities has long been a goal of modern biology. The recently discovered Type IIL MmeI family of restriction-and-modification (RM) enzymes that possess a shared target recognition domain provides a framework for engineering such new specificities. However, a lack of structural information on Type IIL enzymes has limited the repertoire that can be rationally engineered. We report here a crystal structure of MmeI in complex with its DNA substrate and an S-adenosylmethionine analog (Sinefungin). The structure uncovers for the first time the interactions that underlie MmeI-DNA recognition and methylation (5'-TCCRAC-3'; R = purine) and provides a molecular basis for changing specificity at four of the six base pairs of the recognition sequence (5'-TCCRAC-3'). Surprisingly, the enzyme is resilient to specificity changes at the first position of the recognition sequence (5'-TCCRAC-3'). Collectively, the structure provides a basis for engineering further derivatives of MmeI and delineates which base pairs of the recognition sequence are more amenable to alterations than others.

PubMed: 27082731
DOI: 10.1371/journal.pbio.1002442

実験手法

X-RAY DIFFRACTION (2.5964 Å)