Crystallization and preliminary crystallographic analysis of hygromycin B phosphotransferase from Escherichia coli.
Aminoglycoside antibiotics, such as hygromycin, kanamycin, neomycin, spectinomycin and streptomycin, inhibit protein synthesis by acting on bacterial and eukaryotic ribosomes. Hygromycin B phosphotransferase (Hph; EC 2.7.1.119) converts hygromycin B to 7''-O-phosphohygromycin using a phosphate moiety from ATP, resulting in the loss of its cell-killing activity. The Hph protein has been crystallized for the first time using a thermostable mutant and the hanging-drop vapour-diffusion method. The crystal provided diffraction data to a resolution of 2.1 A and belongs to space group P3(2)21, with unit-cell parameters a = b = 71.0, c = 125.0 A. Crystals of complexes of Hph with hygromycin B and AMP-PNP or ADP have also been obtained in the same crystal form as that of the apoprotein.
Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502, Japan.
Daisuke
D
Iino
Yasuaki
Y
Takakura
Mika
M
Kuroiwa
Ryouta
R
Kawakami
Yasuyuki
Y
Sasaki
Takayuki
T
Hoshino
Kanju
K
Ohsawa
Akira
A
Nakamura
Shunsuke
S
Yajima
D002934:Cinnamates / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000737:chemistry / Q000378:metabolism
D002934:Cinnamates; D029968:Escherichia coli Proteins; D011500:Protein Synthesis Inhibitors; D006921:Hygromycin B; C026273:hygromycin A; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase