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Inhibition of Bacillus cereus spore outgrowth by covalent modification of a sulfhydryl group by nitrosothiol and iodoacetate.

ABSTRACT

Nitrosothiols with the general structure RSN==O were studied as a model system of bacteriostatic action toward outgrowing bacterial spores. With a Taft plot analysis, the influence of the structure of the R group on the inhibitory effectiveness of a series of nitrosothiols showed that effectiveness as an inhibitor of Bacillus cereus T outgrowth correlated with the electron withdrawal of R, but that size, shape, charge, hydrophobicity, and transportability had little influence. This was interpreted to mean that nitrosothiols do not traverse the membrane to act. The Taft plot together with competition data between nitrosothiol and iodoacetate indicated that the mode of nitrosothiol action is the covalent modification of a sulfhydryl group, probably to form RSN(OH)--SX, where --SX is derived from a sensitive spore sulfhydryl group. Cooperativity effects indicated that outgrowth inhibition is accompanied by a conformational change occurring upon sulfhydryl group modification, which is communicated among at least three to five subunits. Uptake of label during spore germination indicated that most of the sulfhydryl groups which can be modified are associated with the inhibitory event. These data suggest that this sulfhydryl group may be sufficiently unique that inhibitors designed to interfere specifically with it could have value as bacteriostatic agents.

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