Identification of a mutation in Escherichia coli F1-ATPase beta-subunit conferring resistance to aurovertin.
A mutation conferring aurovertin resistance on Escherichia coli F1-ATPase was identified as R398----H in the F1 beta-subunit. Beta-subunit from the mutant does not bind aurovertin; therefore our results suggest the region of sequence around residue beta-398 is involved in aurovertin binding. Since nucleotide and aurovertin binding to isolated beta-subunit are not mutually exclusive, the data further suggest that the beta-subunit catalytic nucleotide-binding domain does not include residue 398. The mutation prevented aurovertin inhibition of ATPase at pH 6 and 8.5, implying charge on the arginine side-chain is not a major determinant of aurovertin binding or that the pK of R398 is shifted due to a peculiar environment. The equivalent residue is usually arginine in F1 beta-subunits of different species; notably in the aurovertin-insensitive thermophilic bacterium PS3 F1-ATPase, this residue is phenylalanine.
Lee
R S
RS
Department of Biochemistry, University of Rochester Medical Center, NY 14642.
Pagan
J
J
Satre
M
M
Vignais
P V
PV
Senior
A E
AE
D001313:Aurovertins / Q000494:pharmacology*; D001665:Binding Sites; D003001:Cloning, Molecular; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D009154:Mutation; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000235:genetics*; D011714:Pyrans / Q000494:pharmacology*
D001313:Aurovertins; D011714:Pyrans; D006180:Proton-Translocating ATPases