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Use of a novel method to find substrates of protein kinase C delta identifies M2 pyruvate kinase.


ABSTRACT: Protein kinase C (PKC) family members have been implicated in numerous cellular processes. However, identifying the substrates of each PKC isozyme remains a challenge. Here, we describe a method using two-dimensional (2D) isoelectric focusing gel electrophoresis to identify substrates of delta PKC (deltaPKC) in MCF-7 breast carcinoma cells. We show that M2 pyruvate kinase is a substrate of deltaPKC, and further characterize the interaction between M2 pyruvate kinase and deltaPKC in MCF-7 cells by immunoprecipitation. deltaPKC activation in vitro or in cells did not appear to alter the enzyme activity or polymerization of M2 pyruvate kinase.

SUBMITTER: Siwko S 

PROVIDER: S-EPMC1931518 | biostudies-literature | 2007

REPOSITORIES: biostudies-literature

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Use of a novel method to find substrates of protein kinase C delta identifies M2 pyruvate kinase.

Siwko Stefan S   Mochly-Rosen Daria D  

The international journal of biochemistry & cell biology 20070124 5


Protein kinase C (PKC) family members have been implicated in numerous cellular processes. However, identifying the substrates of each PKC isozyme remains a challenge. Here, we describe a method using two-dimensional (2D) isoelectric focusing gel electrophoresis to identify substrates of delta PKC (deltaPKC) in MCF-7 breast carcinoma cells. We show that M2 pyruvate kinase is a substrate of deltaPKC, and further characterize the interaction between M2 pyruvate kinase and deltaPKC in MCF-7 cells b  ...[more]

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