JBC, Vol. 250, Issue 19, 7739-7746, Oct, 1975
Use of diazido ethidium bromide as a specific probe for mitochondrial functions
R. N. Bastos
The diazido derivative of ethidium bromide has been synthesized as a
potential photoaffinity label and shown to be at least as effective as a
mitochondrial mutagen as the parent compound, with a similar mode of
action. Exposure of mitochondria of Saccharomyces cerevisiae to the
compound, followed by ultraviolet-irradiation, which converts it to the
highly reactive dinitrene, results in its specific binding to a single
component which has been tentatively identified as the smallest polypeptide
(subunit 9) of the membrane-bound ATPase. An analogus reaction is also
obtained with the soluble, oligomycin-sensitive ATPase complex but not with
the F1-ATPase itself. The reaction with the ATPase complex can also be
monitored by fluorescence enhancement and by this attribute, as well as by
other criteria, diazido-ethidium bromide, ethidium bromide itself,
euflavine, N,N'-dicyclohexylcarbodiimide, 2,4-dinitrophenol, and
2-azido-4-nitrophenol all appear to compete for the same, lipophilic,
binding site. A mitochondrial mutation (73/1) (see Flury, U., Feldman, F.,
and Mahler, H.R. (1974) J. Biol. Chem. 249, 6630-6637) produces a
photoaffinity product with an altered electrophoretic mobility and
molecular weight.
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