Crystallization and preliminary X-ray diffraction analysis of the full-length Mg2+ transporter MgtE.
The MgtE family of Mg(2+) transporters are ubiquitously conserved in all three domains. The genes encoding full-length MgtE from seven different species were cloned. Three of the seven MgtE transporters were overexpressed and purified for use in crystallization trials. Only Thermus thermophilus MgtE was successfully crystallized using the sitting-drop vapour-diffusion method. Selenomethionine-substituted (SeMet) crystals were obtained by cross-microseeding using the native microcrystals. The SeMet crystals diffracted X-rays to 3.5 A resolution using synchrotron radiation and belong to space group C222(1), with unit-cell parameters a = 118.3, b = 134.9, c = 366.2 A. Structure determination is in progress.
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.
Motoyuki
M
Hattori
Yoshiki
Y
Tanaka
Shuya
S
Fukai
Ryuichiro
R
Ishitani
Osamu
O
Nureki
D017920:Antiporters / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D002413:Cations, Divalent; D003460:Crystallization; D008274:Magnesium / Q000737:chemistry* / Q000378:metabolism; D016963:Thermus thermophilus; D014961:X-Ray Diffraction / Q000379:methods
D017920:Antiporters; D001426:Bacterial Proteins; D002413:Cations, Divalent; C092192:MgtE protein, bacteria; D008274:Magnesium