Crystallization and preliminary X-ray analysis of the tRNA thiolation enzyme MnmA from Escherichia coli complexed with tRNA(Glu).
MnmA catalyzes a sulfuration reaction to synthesize 2-thiouridine at the wobble positions of tRNA(Glu), tRNA(Gln) and tRNA(Lys) in Escherichia coli. The binary complex of MnmA and tRNA(Glu) was crystallized in two different crystal forms: forms I and II. Cocrystallization of MnmA-tRNA(Glu) with ATP yielded form III crystals. The three crystal forms diffracted to 3.1, 3.4 and 3.4 angstroms resolution, respectively, using synchrotron radiation at SPring-8. These crystals belong to space groups C2, I2(1)2(1)2(1) and C2, with unit-cell parameters a = 225.4, b = 175.8, c = 53.0 angstroms, beta = 101.6 degrees, a = 101.5, b = 108.0, c = 211.2 A and a = 238.1, b = 102.1, c = 108.2 angstroms, beta = 117.0 degrees, respectively. The asymmetric units of these crystals are expected to contain two, one and two MnmA-tRNA(Glu) complexes, respectively.
Numata
Tomoyuki
T
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama-shi, Kanagawa 226-8501, Japan.
Ikeuchi
Yoshiho
Y
Fukai
Shuya
S
Adachi
Hiroaki
H
Matsumura
Hiroyoshi
H
Takano
Kazufumi
K
Murakami
Satoshi
S
Inoue
Tsuyoshi
T
Mori
Yusuke
Y
Sasaki
Takatomo
T
Suzuki
Tsutomu
T
Nureki
Osamu
O
D003001:Cloning, Molecular; D003460:Crystallization; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012352:RNA, Transfer, Glu / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014961:X-Ray Diffraction
D029968:Escherichia coli Proteins; D012352:RNA, Transfer, Glu; D011994:Recombinant Proteins; C510958:MnmA protein, E coli