title: Analysis of CaM-kinase signaling in cells.
abstract: A change in intracellular free calcium is a common signaling mechanism that modulates a wide array of physiological processes in most cells. Responses to increased intracellular Ca(2+) are often mediated by the ubiquitous protein calmodulin (CaM) that upon binding Ca(2+) can interact with and alter the functionality of numerous proteins including a family of protein kinases referred to as CaM-kinases (CaMKs). Of particular interest are multifunctional CaMKs, such as CaMKI, CaMKII, CaMKIV and CaMKK, that can phosphorylate multiple downstream targets. This review will outline several protocols we have used to identify which members and/or isoforms of this CaMK family mediate specific cellular responses with a focus on studies in neurons. Many previous studies have relied on a single approach such as pharmacological inhibitors or transfected dominant-negative kinase constructs. Since each of these protocols has its limitations, that will be discussed, we emphasize the necessity to use multiple, independent approaches in mapping out cellular signaling pathways.
authors.affiliation: Program in Neuroscience, Department of Veterinary and Comparative Anatomy, Pharmacology and Physiology, Washington State University, Pullman, WA, USA. waymang@vetmed.wsu.edu
authors.forename: Gary A
authors.initials: GA
authors.lastname: Wayman
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tokumitsu
authors.forename: Monika A
authors.initials: MA
authors.lastname: Davare
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Soderling
mesh_terms: D002118:Calcium / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering; D015398:Signal Transduction
chemical_list: D047428:Protein Kinase Inhibitors; D034741:RNA, Small Interfering; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium