19F NMR investigation of F(1)-ATPase of Escherichia coli using fluorotryptophan labeling.
Growth of Escherichia coli in the presence of glyphosate, an inhibitor of aromatic amino acid biosynthesis, has permitted the production of proton-dislocating ATPase that is specifically labeled with 5-fluorotryptophan. Five sets of (19)F resonances could be assigned to each tryptophan residue by lauryldimethylamine oxide and carboxypeptidase treatment. On labeling with 4-chloro-7-nitro-benzofurazan, the label attached to b155Lys, which is known to be in the catalytic site, which caused one of the residues, b108Trp, to become nonequivalent. (19)F NMR spectroscopic investigation of internally fluorotryptophan-labeled F(1)-ATPase will provide valuable information about the asymmetric nature of F(1)-ATPase and the conformational changes induced by ligand binding.
Lee
H W
HW
Department of Biochemistry and Institute of Basic Medical Science, Yonsei University Wonju College of Medicine, Wonju 220-701, Korea.
Sohn
J H
JH
Yeh
B I
BI
Choi
J W
JW
Jung
S
S
Kim
H W
HW
D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry; D002268:Carboxypeptidases / Q000737:chemistry; D004123:Dimethylamines / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology*; D005461:Fluorine; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D006180:Proton-Translocating ATPases / Q000096:biosynthesis / Q000378:metabolism*; D014364:Tryptophan / Q000037:antagonists & inhibitors / Q000378:metabolism; D000097797:Glyphosate
D009327:4-Chloro-7-nitrobenzofurazan; D002268:Carboxypeptidases; D004123:Dimethylamines; D005461:Fluorine; D005998:Glycine; D006180:Proton-Translocating ATPases; D014364:Tryptophan; D000097797:Glyphosate; C014518:dodecyldimethylamine oxide