Crystallization and preliminary X-ray diffraction analysis of MspI restriction endonuclease in complex with its cognate DNA.

Autor: O'Loughlin, Terence J., Xu, Qian, Kucera, Rebecca B., Dorner, Lydia F., Sweeney, Suzanne, Schildkraut, Ira, Guo, Hwai-Chen
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Zdroj: Acta Crystallographica: Section D (Wiley-Blackwell); Dec2000, Vol. 56 Issue 12, p1652-1655, 4p
Abstrakt: The MspI restriction endonuclease is a type II restriction enzyme. Unlike all other restriction enzymes with known structures, MspI recognizes the palindromic tetranucleotide sequence 5′-C/CGG and cleaves it as indicated by the `I' to produce DNA products with 5' two- base overhangs. Owing to the nature of its cleavage pattern, it is likely that MspI would represent a new structural class of restriction endonucleases. Crystals of the dimeric MspI restriction enzyme bound to a duplex DNA molecule containing the specific recognition sequence have been obtained by vapor-diffusion techniques in the presence of polyethylene glycol as precipitant. The crystals belong to the monoclinic space group P21, with unit-cell parameters a 50.2, b = 131.6, c = 59.3 Å, β = 109.7°. The crystals contain one dimeric complex in the asymmetric unit. A complete native data set has been collected to a resolution of 2.05 A by cryo-crystallographic methods, with an Rmerge of 4.0%. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index