Calmodulin purification

Removal of contaminating proteins (which bind) during calmodulin purification Purification of pancreatic carboxypeptidase Purification of crayfish carboxypeptidase Purification of wheat-germ aspartate trans-carbamoylase... 

Ballihaut, G., Tastet, L., Pecheyran, C., Bouyssiere, B., Donard, O., Grimaud, R., and Lobinski, R., Biosynthesis, purification and analysis of selenomethionyl calmodulin by gel electrophoresis-laser ablation-ICP-MS and capillary HPLC-ICP-MS peptide mapping following in-gel tryptic digestion. Journal of Analytical Atomic Spectrometry 20(6), 493 99, 2005. 

Mayer, B., John, M., and Bohme, E. (1990a). Purification of aCa /calmodulin-dependent nitric oxide synthase from porcine cerebellum. FEBS Lett. 277, 215-219. 

Walsh, M.P. Vallet, B. Autric, E Demaille, J.G. Purification and characterization of bovine cardiac calmodulin-dependent myosin light chain kinase. J. Biol. Chem., 254, 12136-12144 (1979)... 

Bartelt, D.C. Moroney, S. Wolff, D.J. Purification, characterization and substrate specificity of calmodulin-dependent myosin light-chain kinase from bovine brain. Biochem. J., 247, 747-756 (1987)... 

Ikebe, M. Reardon, S. Scott-Woo, G.C. Zhou, Z. Koda, Y. Purification and characterization of calmodulin-dependent multifunctional protein kinase from smooth muscle isolation of caldesmon kinase. Biochemistry, 29, 11242-11248 (1990)... 

Rieker, J.P. Swanljung-Collins, H. Montibeller, J. Collins, J.H. Purification and characterization of a calmodulin-dependent myosin heavy chain kinase from intestinal brush border. J. Biol. Chem., 262, 15262-15268 (1987)... 

Edelman, A. M., Mitchelhill, K. I., Seibert, M. A., Anderson, K. A., Hook, S. S., Stapleton, D., Goldstein, E. G., Means, A. R. and Kemp, B. E., 1996, Multiple Ca2+ -calmodulin-dependentprotein kinase kinases from rat brain Purification, regulation by Ca2+ -calmodulin and partial amino acid sequence, The Journal of Biological Chemistry, 271, pp 10806-10810. 

Figure 3 Silver stained SDS-PAGE gel of iPLA2-VI purification. Lane 1, molecular mass markers lane 2,20 pi of pool from octyl-Sepharose column lane 3,20 pi of fraction 28 from the Calmodulin-agarose column lane 4,20 pi of fraction 29 from the Calmodulin-agarose column and lane 5,20 pi of fraction 30 from the Calmodulin-agarose column. (From Ref. 41.)...

Douglas, P., Moorhead, G., Hong, Y., Morrice, N., and MacKintosh, C., 1998, Purification of a nitrate reductase kinase from Spinaceaa oleracea leaves, and its identification as a calmodulin-domain protein kinase, Planta 206 43511442. 

We thank Dr. K. Beckingham and colleagues (Rice University, Texas, USA) for providing the mutant calmodulins and Peter Fletcher (N.I.M.R.) for synthesis and purification of the peptides. 

Sobue, K., Muramoto, K., Fujita, M. and Kakiuchi, S. (1981). Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin. Proc. Natl. Acad. Sci. USA 78, 5652-5655. 

In brain, a large number of particulate soluble proteins are phosphory-lated in the presence of Mg " -ATP and Ca +—calmodulin. These phosphorylations can be blocked by phenothiazines and do not occur in the presence of cAMP or cGMP. Furthermore, they are unaltered by the Walsh inhibitor, which is specific for the cAMP catalytic subunit. Hence, investigators have concluded that there is present in both particulate and soluble fractions of brain a Ca " -calmodulin-dependent protein kinase that is insensitive to cyclic nucleotides and that contains its own set of protein substrates. A few reports have appeared in the literature claiming purification of a calcium-calmodulin-dependent kinase from mammalian brain. The en-... 

Sobieszek A, Babiychuk EB, Ortner B, Borkowski J (1997a) Purification and characterization of a kinase-associated myofibriOar smooth muscle myosin light chain phosphatase possessing a calmodulin-targeting subunit. J Biol Chem 272 7027-7033... 

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