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Purification and properties of a multifunctional calcium/calmodulin-dependent protein kinase from rat pancreas.

Publication ,  Journal Article
Cohn, JA; Kinder, B; Jamieson, JD; Delahunt, NG; Gorelick, FS
Published in: Biochim Biophys Acta
May 18, 1987

A calcium/calmodulin-dependent protein kinase (Ca/calmodulin protein kinase) was purified from rat pancreas using hydrophobic chromatography followed by gel filtration and affinity chromatography. Ca/calmodulin protein kinase from pancreas resembled previously described multifunctional Ca/calmodulin protein kinases from other tissues with respect to substrate specificity, autophosphorylation on serine and threonine residues, and catalytic and hydrodynamic properties. While Ca/calmodulin protein kinase from other tissues contains subunits of 53-60 kDa with variable proportions of a smaller 50-52 kDa subunit, pancreatic Ca/calmodulin protein kinase was found to contain a single component of 51 kDa. Experiments mixing brain Ca/calmodulin protein kinase with pancreatic homogenate suggest that the absence of a larger subunit in the pancreatic Ca/calmodulin protein kinase is not due to proteolytic degradation during enzyme preparation. Ca/calmodulin protein kinase binding to 125I-labeled calmodulin in solution was demonstrated using the photoaffinity cross-linker, N-hydroxysuccinimidyl-4-azidobenzoate. 125I-labeled calmodulin binding to Ca/calmodulin protein kinase was also demonstrated using filters containing Ca/calmodulin protein kinase transferred from polyacrylamide gels after two-dimensional gel electrophoresis. Finally, the ribosomal substrate for Ca/calmodulin protein kinase was identified as the ribosomal protein, S6. The purification procedure presented in this study promises to be useful in characterizing Ca/calmodulin protein kinase in other tissues and in clarifying the role of these enzymes in cellular function.

Duke Scholars

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

May 18, 1987

Volume

928

Issue

3

Start / End Page

320 / 331

Location

Netherlands

Related Subject Headings

  • Synapsins
  • Substrate Specificity
  • Rats
  • Protein Kinases
  • Protein Conformation
  • Photochemistry
  • Phosphorylation
  • Pancreas
  • Nerve Tissue Proteins
  • Macromolecular Substances
 

Citation

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Cohn, J. A., Kinder, B., Jamieson, J. D., Delahunt, N. G., & Gorelick, F. S. (1987). Purification and properties of a multifunctional calcium/calmodulin-dependent protein kinase from rat pancreas. Biochim Biophys Acta, 928(3), 320–331. https://doi.org/10.1016/0167-4889(87)90192-3
Cohn, J. A., B. Kinder, J. D. Jamieson, N. G. Delahunt, and F. S. Gorelick. “Purification and properties of a multifunctional calcium/calmodulin-dependent protein kinase from rat pancreas.Biochim Biophys Acta 928, no. 3 (May 18, 1987): 320–31. https://doi.org/10.1016/0167-4889(87)90192-3.
Cohn JA, Kinder B, Jamieson JD, Delahunt NG, Gorelick FS. Purification and properties of a multifunctional calcium/calmodulin-dependent protein kinase from rat pancreas. Biochim Biophys Acta. 1987 May 18;928(3):320–31.
Cohn, J. A., et al. “Purification and properties of a multifunctional calcium/calmodulin-dependent protein kinase from rat pancreas.Biochim Biophys Acta, vol. 928, no. 3, May 1987, pp. 320–31. Pubmed, doi:10.1016/0167-4889(87)90192-3.
Cohn JA, Kinder B, Jamieson JD, Delahunt NG, Gorelick FS. Purification and properties of a multifunctional calcium/calmodulin-dependent protein kinase from rat pancreas. Biochim Biophys Acta. 1987 May 18;928(3):320–331.

Published In

Biochim Biophys Acta

DOI

ISSN

0006-3002

Publication Date

May 18, 1987

Volume

928

Issue

3

Start / End Page

320 / 331

Location

Netherlands

Related Subject Headings

  • Synapsins
  • Substrate Specificity
  • Rats
  • Protein Kinases
  • Protein Conformation
  • Photochemistry
  • Phosphorylation
  • Pancreas
  • Nerve Tissue Proteins
  • Macromolecular Substances