The mechanism of activation of protein kinase C isoforms by filamentous actin (F-actin) wasinvestigated with respect to isozyme specificity and phorbol ester and Ca
2+ dependencies. It was foundthat the "conventional" (cPKC),
![](/images/gifchars/alpha.gif)
,
![](/images/gifchars/beta2.gif)
I,
![](/images/gifchars/beta2.gif)
II, and
![](/images/gifchars/gamma.gif)
, "novel" (nPKC)
![](/images/gifchars/delta.gif)
and
![](/images/gifchars/epsilon.gif)
, and "atypical" (aPKC)
![](/images/gifchars/zeta.gif)
isoforms were each activated by F-actin with varying potencies. The level of activity along with theaffinity for binding to F-actin was further potentiated by the phorbol ester
4![](/images/gifchars/beta2.gif)
-12-
O-tetradecanoylphorbol13-acetate (TPA), the potency of which again varied for each isoform. By contrast to the other cPKCisoforms, the level of cPKC-
![](/images/gifchars/gamma.gif)
activity was unaffected by TPA, as was also the case for aPKC-
![](/images/gifchars/zeta.gif)
. It wasfound that whereas in the absence of F-actin the soluble form of cPKC-
![](/images/gifchars/beta2.gif)
I contained two phorbol esterbinding sites of low and high affinity, respectively, as previously reported for cPKC-
![](/images/gifchars/alpha.gif)
[
Slater et al. (1998)
J. Biol. Chem. 273, 23160-23168], the F-actin-bound form of the isozyme contained only a single siteof relatively low affinity. The level of TPA required to induce cPKC-
![](/images/gifchars/alpha.gif)
, -
![](/images/gifchars/beta2.gif)
I, and -
![](/images/gifchars/beta2.gif)
II activity and thebinding of these isozymes to F-actin was reduced in the presence of Ca
2+. By contrast, the activity ofcPKC-
![](/images/gifchars/gamma.gif)
was unaffected by Ca
2+, as were the activities of nPKC-
![](/images/gifchars/delta.gif)
and -
![](/images/gifchars/epsilon.gif)
and aPKC-
, as expected.Thus, the interaction with F-actin appears to be a general property of each of the seven PKC isozymestested. However, isoform specificity may, in part, be directed by differences in the phorbol ester andCa
2+ dependences, which, with the notable exception of cPKC-
, appear to resemble those observed forthe activation of each isoform by membrane association. The observation that cPKC isoforms maytranslocate to F-actin as well as the membrane as a response to an elevation of Ca
2+ levels may allow forthe functional coupling of fluctuations of intracellular Ca
2+ levels through cPKC to F-actin cytoskeleton-mediated processes.