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JBC, Vol. 251, Issue 21, 6646-6652, Nov, 1976
J. A. Evans and M. P. Deutscher
We have examined the cation requirements of rabbit liver tRNA
nucleotidyltransferase. The enzyme had an absolute requirement fo a
divalent cation which could be satisfied by Mg2+, Mn2+ or Co2+. In contrast
to the Escherichia coli enzyme, we have found no evidence to implicate Zn2+
in the action of rabbit liver tRNA nucleotidyltransferase. We have also
identified a second cation requirement which was satisfied by divalent or
monovalent cation or polyamines. The polyamines, spermine and spermidine,
were most effective leading to 3-fold higher rates of nucleotide
incorporation than could be obtained at any concentration of the other
cations. However, the polyamines were not an absolute requirement for
enzyme activity. The polyamine stimulation of tRNA nucleotidyltransferase
was not due to alteration of the pH or ionic strength of the reaction
mixture or reactivation of denatured tRNA or enzyme. Polyamines also had no
effect on the apparent Km values of the substrates. Spermine increased the
specificity of the enzyme for tRNA substrates and also inhibited the
reverse action. Our results suggest that polyamines may be the normal
counterions for tRNA in vivo, and that they affect the rate-limiting step
in tRNA nucleotidyltransferase catalysis.
Polyamine stimulation and cation requirements of rabbit liver tRNA nucleotidyltransferase
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