N,N'-dicyclohexylcarbodiimide and 4-chloro-7-nitrobenzofurazan bind to different beta subunits of the F1 ATPase of Escherichia coli.
The fluorescent thiol reagent 2-(4'-iodoacetamidoanilino)naphthalene-6-sulfonic acid (IAANS) labels the gamma, delta, and one of the three beta subunits of the F1 ATPase from Escherichia coli (ECF1). This is the same beta subunit which incorporates 4-chloro-7-nitrobenzofurazan (Nbf) [H. Stan-Lotter and P. D. Bragg (1986) Eur. J. Biochem. 154, 321-327]. After inactivation of ECF1 with N,N'-dicyclohexylcarbodiimide (DCCD), IAANS labels in addition to the beta, gamma, and delta subunits also the alpha subunit. This suggests a conformational change of ECF1 upon binding of DCCD. The beta subunit which incorporates DCCD does does not bind IAANS. Likewise, IAANS-modified ECF1 does not incorporate DCCD into the same beta subunit. It is concluded that DCCD and Nbf bind to different beta subunits. Since neither of these reagents binds to that beta subunit which can be crosslinked to to the epsilon subunit by 1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide, these data show that there is a difference in the chemical reactivity of each of the three beta subunits of ECF1, despite their identical primary structures. This suggests that there is an asymmetry in the F1 molecule.
Stan-Lotter
H
H
Bragg
P D
PD
D009327:4-Chloro-7-nitrobenzofurazan / Q000494:pharmacology*; D002234:Carbodiimides / Q000378:metabolism*; D004024:Dicyclohexylcarbodiimide / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology*; D005456:Fluorescent Dyes; D007525:Isoelectric Focusing; D009282:Naphthalenesulfonates / Q000378:metabolism; D010069:Oxadiazoles / Q000494:pharmacology*; D006180:Proton-Translocating ATPases / Q000378:metabolism*
D002234:Carbodiimides; D005456:Fluorescent Dyes; D009282:Naphthalenesulfonates; D010069:Oxadiazoles; D004024:Dicyclohexylcarbodiimide; C011488:IAANS; D006180:Proton-Translocating ATPases; D009327:4-Chloro-7-nitrobenzofurazan