Crystallization and preliminary X-ray diffraction analysis of MspI restriction endonuclease in complex with its cognate DNA.
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 '/' 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 P2(1), with unit-cell parameters a = 50.2, b = 131.6, c = 59.3 A, beta = 109.7 degrees. 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 R(merge) of 4.0%.
O'Loughlin
T J
TJ
Department of Physiology and Structural Biology, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118-2526, USA.
Xu
Q
Q
Kucera
R B
RB
Dorner
L F
LF
Sweeney
S
S
Schildkraut
I
I
Guo
H C
HC
D003460:Crystallization; D004247:DNA / Q000737:chemistry*; D019003:Deoxyribonuclease HpaII / Q000737:chemistry* / Q000302:isolation & purification; D009016:Moraxella / Q000201:enzymology; D011487:Protein Conformation; D014961:X-Ray Diffraction
D004247:DNA; D019003:Deoxyribonuclease HpaII