Lopez-Garcia Laura A, Schulze Jörg O, Fröhner Wolfgang, Zhang Hua, Süss Evelyn, Weber Nadja, Navratil Jeanette, Amon Sabine, Hindie Valerie, Zeuzem Stefan, Jørgensen Thomas J D, Alzari Pedro M, Neimanis Sonja, Engel Matthias, Biondi Ricardo M
Research Group PhosphoSites, Department of Internal Medicine I, Universitätsklinikum Frankfurt, 60590 Frankfurt, Germany.
Chem Biol. 2011 Nov 23;18(11):1463-73. doi: 10.1016/j.chembiol.2011.08.010.
Protein kinases are key mediators of cellular signaling, and therefore, their activities are tightly controlled. AGC kinases are regulated by phosphorylation and by N- and C-terminal regions. Here, we studied the molecular mechanism of inhibition of atypical PKCζ and found that the inhibition by the N-terminal region cannot be explained by a simple pseudosubstrate inhibitory mechanism. Notably, we found that the C1 domain allosterically inhibits PKCζ activity and verified an allosteric communication between the PIF-pocket of atypical PKCs and the binding site of the C1 domain. Finally, we developed low-molecular-weight compounds that bind to the PIF-pocket and allosterically inhibit PKCζ activity. This work establishes a central role for the PIF-pocket on the regulation of PKCζ and allows us to envisage development of drugs targeting the PIF-pocket that can either activate or inhibit AGC kinases.
蛋白激酶是细胞信号传导的关键介质，因此，它们的活性受到严格控制。AGC激酶受磷酸化以及N端和C端区域的调节。在此，我们研究了非典型PKCζ的抑制分子机制，发现N端区域的抑制不能用简单的假底物抑制机制来解释。值得注意的是，我们发现C1结构域通过变构抑制PKCζ活性，并证实了非典型PKC的PIF口袋与C1结构域结合位点之间的变构通讯。最后，我们开发了与PIF口袋结合并变构抑制PKCζ活性的低分子量化合物。这项工作确立了PIF口袋在PKCζ调节中的核心作用，并使我们能够设想开发靶向PIF口袋的药物，这些药物可以激活或抑制AGC激酶。
