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Effect of thiourea and substituted thioureas on dynein ATPase and on the turbidity response of tetrahymena cilia
Authors:J J Blum  A Hayes
Abstract:The effects of thioura and of several substituted thioureas–phenylthiourea, α-naphtylthiourea, metiamide, and burimamide–on dynein ATPase have been studied. The substituted thioureas are over 30 times more potent than thiourea in causing enhancement of 30S dynein ATPase activity and inhibition of 14S dynein ATPase activity. The effects of thiourea and phenylthiourea can be prevented by very low concentrations of β-mercaptoethanol or dithiotheritol. Axonemal ATPase is also enhanced by the thioureas, but the reaction proceeds more slowly than for solubilized 30S dynein. Enhancement of 30S dynein ATPase by metiamide is prevented by low (~ 1 μM) concentrations of ATP and, less effectively, by AMP-PNP, but not by AMP-PCP even though the latter is a stronger inhibitor of 30S dynein ATPase than is AMP-PNP. The thioureas inhibit the ATP-induced decrease in turbidity (measured as ΔA350) of axonemal suspensions. Inhibition of the turbidity response is also prevented by low concentrations of β-mercaptoethanol, but, in contrast to the irreversible enhancement of ATPase activity, inhibition of the turbidity response is largely reversible. The ability of 30S dynein to rebind onto twice extracted axonemes is not changed by treatment with phenylthiourea or metiamide. These observations indicate that the thioureas react with at least two sets of SH or S–S groups on axonemes. Reaction with the group(s) on the 30S dynein causes an apparently irreversible enhancement of ATPase activity. Reaction with another group(s) causes a reversible inhibition of the turbidity response.
Keywords:thiourea  cilia  dyneins  ATPase  sulfhydryl groups  turbidity response
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