Site-directed mutagenesis and preliminary x-ray crystallographic studies of the tabtoxin resistance protein.
Tabtoxin resistance protein (TTR) is an enzyme that catalyzes the acetylation of tabtoxin rendering tabtoxin-producing pathogens tolerant to their own phytotoxins. According to the structure based detoxification mechanism of TTR, three site-directed mutants Y141F, D130N and Y141F-D130N were constructed and overexpressed in E. coli. The products were then purified and their properties were analyzed by CD and DLS. Crystallization trials of two mutants Y141F andY141F-D130N were preformed.
Ding
Yi
Y
Laboratory of Structural Biology, School of Life Science & Engineering, Tsinghua University, Beijing 100084, China.
Li
Shentao
S
Li
Xiaofeng
X
Sun
Fei
F
Liu
Jinyuan
J
Zhao
Nanming
N
Rao
Zihe
Z
D000107:Acetylation; D000123:Acetyltransferases / Q000737:chemistry* / Q000235:genetics*; D001426:Bacterial Proteins; D002942:Circular Dichroism; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008658:Inactivation, Metabolic; D007526:Isoelectric Point; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics*; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000123:Acetyltransferases; C441636:tabtoxin-resistance protein, Pseudomonas syringae