2GIJ

Q138F HincII bound to cognate DNA GTTAAC and Ca2+

2GIJ の概要

• エントリーDOI: 10.2210/pdb2gij/pdb
• 関連するPDBエントリー: 1TX3 2GIE 2GIG 2GIH 2GII
• 分子名称: 5'-D(*GP*CP*CP*GP*GP*TP*TP*AP*AP*CP*CP*GP*GP*C)-3', Type II restriction enzyme HincII, SODIUM ION, ... (5 entities in total)
• 機能のキーワード: protein dna complex, indirect readout, dna intercalation, endonuclease, hydrolase-dna complex, hydrolase/dna
• 由来する生物種: Haemophilus influenzae
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 68344.92

構造登録者

Horton, N.C.,Joshi, H.K. (登録日: 2006-03-28, 公開日: 2006-07-18, 最終更新日: 2024-02-14)

主引用文献

Joshi, H.K.,Etzkorn, C.,Chatwell, L.,Bitinaite, J.,Horton, N.C.
Alteration of Sequence Specificity of the Type II Restriction Endonuclease HincII through an Indirect Readout Mechanism.
J.Biol.Chem., 281:23852-23869, 2006

PubMed Abstract: The functional and structural consequences of a mutation of the DNA intercalating residue of HincII, Q138F, are presented. Modeling has suggested that the DNA intercalation by Gln-138 results in DNA distortions potentially used by HincII in indirect readout of its cognate DNA, GTYRAC (Y = C or T, R = A or G) (Horton, N. C., Dorner, L. F., and Perona, J. J. (2002) Nat. Struct. Biol. 9, 42-47). Kinetic data presented here indicate that the mutation of glutamine 138 to phenylalanine (Q138F) results in a change in sequence specificity at the center two base pairs of the cognate recognition site. We show that the preference of HincII for cutting, but not binding, the three cognate sites differing in the center two base pairs has been altered by the mutation Q138F. Five new crystal structures are presented including Q138F HincII bound to GTTAAC and GTCGAC both with and without Ca2+ as well as the structure of wild type HincII bound to GTTAAC. The Q138F HincII/DNA structures show conformational changes in the protein, bound DNA, and at the protein-DNA interface, consistent with the formation of adaptive complexes. Analysis of these structures and the effect of Ca2+ binding on the protein-DNA interface illuminates the origin of the altered specificity by the mutation Q138F in the HincII enzyme.

PubMed: 16675462
DOI: 10.1074/jbc.M512339200

実験手法

X-RAY DIFFRACTION (1.93 Å)