1FOK

STRUCTURE OF RESTRICTION ENDONUCLEASE FOKI BOUND TO DNA

1FOK の概要

• エントリーDOI: 10.2210/pdb1fok/pdb
• 分子名称: DNA (5'-D(*TP*CP*GP*GP*AP*TP*GP*AP*TP*AP*AP*CP*GP*CP*TP*AP*G P*TP*CP*A)-3'), DNA (5'-D(*AP*TP*GP*AP*CP*TP*AP*GP*CP*GP*TP*TP*AP*TP*CP*AP*T P*CP*CP*G)-3'), PROTEIN (FOKI RESTRICTION ENDONUCLEASE), ... (4 entities in total)
• 機能のキーワード: complex (endonuclease-dna), type iis, restriction endonuclease, deoxyribonuclease, dna hydrolysis, dna cleavage, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Planomicrobium okeanokoites; 詳細
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 77786.20

構造登録者

Aggarwal, D.A.,Wah, J.A.,Hirsch, L.F.,Dorner, I.,Schildkraut, A.K. (登録日: 1997-04-18, 公開日: 1997-12-03, 最終更新日: 2024-02-07)

主引用文献

Wah, D.A.,Hirsch, J.A.,Dorner, L.F.,Schildkraut, I.,Aggarwal, A.K.
Structure of the multimodular endonuclease FokI bound to DNA.
Nature, 388:97-100, 1997

PubMed Abstract: FokI is a member of an unusual class of bipartite restriction enzymes that recognize a specific DNA sequence and cleave DNA nonspecifically a short distance away from that sequence. Because of its unusual bipartite nature, FokI has been used to create artificial enzymes with new specificities. We have determined the crystal structure at 2.8A resolution of the complete FokI enzyme bound to DNA. As anticipated, the enzyme contains amino- and carboxy-terminal domains corresponding to the DNA-recognition and cleavage functions, respectively. The recognition domain is made of three smaller subdomains (D1, D2 and D3) which are evolutionarily related to the helix-turn-helix-containing DNA-binding domain of the catabolite gene activator protein CAP. The CAP core has been extensively embellished in the first two subdomains, whereas in the third subdomain it has been co-opted for protein-protein interactions. Surprisingly, the cleavage domain contains only a single catalytic centre, raising the question of how monomeric FokI manages to cleave both DNA strands. Unexpectedly, the cleavage domain is sequestered in a 'piggyback' fashion by the recognition domain. The structure suggests a new mechanism for nuclease activation and provides a framework for the design of chimaeric enzymes with altered specificities.

PubMed: 9214510
DOI: 10.1038/40446

実験手法

X-RAY DIFFRACTION (2.8 Å)