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Conformational change in beef‐heart mitochondrial F 1 ATPase to ATP synthesis mode induced by dimethylsulfoxide and ATP revealed by sulfhydryl group labeling
Author(s) -
Beharry Seelochan,
Bragg Philip D.
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80975-5
Subject(s) - dtnb , guanidine , chemistry , protein subunit , hydrochloride , dissociation (chemistry) , atpase , biochemistry , adenosine triphosphate , reagent , stereochemistry , enzyme , glutathione , organic chemistry , gene
Treatment of beef‐heart mitochondrial F 1 ATPase with 5,5′‐dithiobis(2‐nitrobenzoic acid) (DTNB) results in the incorporation of 1 mol DTNB/mol F 1 without loss of ATPase activity. Incorporation is not prevented by ATP. Labeling occurs predominantly on an α‐subunit, but also with a significant degree of modification of γ‐ and ε‐subunits. It is suggested that the modified sulfhydryl groups of the α‐, γ‐ and ε‐subunits are in proximity so that only one can be modified by the reagent. Guanidine hydrochloride (0.3 M) dissociates F 1 into its subunits. Eight sulfhydryl groups/mol F 1 can be modified under these conditions. Guanidine hydrochloride does not cause dissociation of F 1 in the presence of 30% (v/v) dimethylsulfoxide (Me 2 SO) and 2 mM ATP. Under these conditions a second molecule of DTNB is incorporated into F 1 with nearly equal modification of the ε‐subunit and an α‐subunit. It is proposed that Me 2 SO and ATP induce a more stable conformation of F 1 , which is resistant to dissociation by guanidine hydrochloride, but in which the site of reaction with DTNB is made more accessible by the guanidine hydrochloride to permit the simultaneous modification of an α‐subunit and the ε‐subunit. Ths conformation is probably that which occurs during ATP synthesis by F 1 in the presence of Me 2 SO.