Protein Kinase C Protocols

Volume 233 of the series Methods in Molecular Biology™ pp 475-489

Genetic Manipulation of Protein Kinase C In Vivo

  • Alex TokerAffiliated withDepartment of Pathology, Beth Israel Deaconess Medical School, Harvard Medical School

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Studies since 1980 have established that protein kinase C (PKC) regulates a plethora of downstream signaling pathways leading to numerous cellular responses. Although much is known concerning PKC regulation by lipid cofactors and phosphorylation, the direct protein substrates that relay the PKC signal remain largely undescribed, although proteins such as myristoylated alanine-rich C kinase (MARCKS) and pleckstrin are well-known examples. Much of the work aimed at defining substrates of PKC has relied on the use of phorbol esters (see Chapter 34) as well as small molecule inhibitors (see Chapter 33). Because of concerns with lack of specificity and cytotoxicity associated with these approaches, new methodologies have been developed to more accurately and specifically manipulate PKC activity in cells and thus tackle the mechanisms by which PKC regulates cell function.