Accession Number : ADA183111
Title : Effects of Hydrazines and Related Compounds on Calcium-Calmodulin Regulated Neuronal Processes.
Descriptive Note : Final rept. 1 Jul 85-1 Feb 87,
Corporate Author : MEDICAL COLL OF VIRGINIA RICHMOND DEPT OF NEUROLOGY
Personal Author(s) : DeLorenzo,Robert J
PDF Url : ADA183111
Report Date : 10 Mar 1987
Pagination or Media Count : 28
Abstract : Results from this investigation have provided a basic understanding of how calcium regulates neuronal cytoskeletal function. Calmodulin-dependent protein kinase activity was investigated in cold stable and cold labile microtubule fractions. Calmodulin-dependent kinase was enriched approximately twenty fold over Cytosol in cold stable microtubule preparations. Calmodulin-dependent Kinase activity in cold stable microtubule preparations phosphorylated microtubule associated protein-2, tubulin, an 80,000 dalton doublet, and several minor phosphoproteins. This endogenous calmodulin-dependent kinase in cold stable microtubule fractions was identical to CaM-Kinase II isolated from rat brain by several criteria including: 1) subunit molecular weights, 2) subunit isoelectric points, 3) calmodulin binding properties, 4) subunit autophophorylation, 5) Calmodulin binding subunit composition, 6)isolation of kinase on calmodulin affinity resin, 7) kinetic parameters, 8) phosphoamino acid phosphorylation sites on tubulin, and, 9) phosphopeptide mapping. Endogenous cold stable calmodulin-dependent kinase activity was isolated from the microtubule fraction by calmodulin affinity resin column chromatography and specifically eluded with EGTA. This kinase fraction contained the calmodulin binding protein and autophosphorylating subunits of CaM Kinase II.
Descriptors : *CALCIUM, *HYDRAZINES, *TOXICITY, *NEUROCHEMISTRY, *PHOSPHORUS TRANSFERASES, ACIDS, PHOSPHORYLATION, SITES, CHEMICAL BONDS, LOW TEMPERATURE, MICROSTRUCTURE, PREPARATION, STABILITY, KINETICS, PHOSPHOPROTEINS, CHROMATOGRAPHY, COLUMN CHROMATOGRAPHY, POLYMERS, NERVE CELLS, BRAIN, RATS, PROTEINS, FIBERS, PEPTIDES, ANTIBODIES
Subject Categories : Toxicology
Distribution Statement : APPROVED FOR PUBLIC RELEASE