Reaction of 2-azido-ATP with beta subunits in the F1-adenosine triphosphatase of Escherichia coli.
The three beta subunits of the Escherichia coli F1-ATPase react independently with chemical reagents (Stan Lotter, H. and Bragg, P.D. (1986) Arch. Biochem. Biophys. 248, 116-120). Thus, one beta subunit is readily cross-linked to the epsilon subunit, another reacts with N,N'-dicyclohexylcarbodiimide (DCCD), and the third one is modified by 4-chloro-7-nitrobenzofurazan (NbfCl). The relationship of the binding site for 2-azido-ATP to the three types of beta subunit recognized by chemical labeling was examined. The binding site for 2-azido-ATP was not associated with a specific type of beta-subunit. There was no relationship between the site of nucleotide and the association of the epsilon subunit with a particular beta subunit. It is concluded that the presence of the epsilon subunit (possibly in association with the other minor subunits) does not determine the position of the catalytic site. The possibility that the lack of a specific relationship between the 2-azido-ATP binding site and a specific beta subunit was due to turnover of the enzyme, making each beta a catalytic site in turn, could not be entirely rejected. However, the rate of hydrolysis of 2-azido-ATP by the DCCD-modified ATPase was very low in the presence of EDTA, and was likely due to catalysis at single sites.
Bragg
P D
PD
Department of Biochemistry, University of British Columbia, Vancouver, Canada.
Hou
C
C
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D001386:Azides / Q000378:metabolism*; D001572:Benzofurans; D001665:Binding Sites; D003432:Cross-Linking Reagents; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D007525:Isoelectric Focusing; D008274:Magnesium / Q000502:physiology; D010777:Photochemistry; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism*
D001386:Azides; D001572:Benzofurans; D003432:Cross-Linking Reagents; D004024:Dicyclohexylcarbodiimide; C042231:2-azidoadenosine 5'-triphosphate; C059027:4-chloro-7-nitrobenzofuran; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D008274:Magnesium