Evidence that casein kinase 2 phosphorylates hepatic microsomal calcium- binding proteins 1 and 2 but not 3

Nengqain Chen, Alan T. Davis, Erhan C. Canbulat, Yuxiu Liu, Said Goueli, Bruce A. McKenzie, Eric D. Eccleston, Khalil Ahmed, Jordan L. Holtzman

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Abstract

We have extensively purified three of the hepatic microsomal intralumenal Ca 2+-binding proteins, CBP1, CBP2, and CBP3, which were originally described by Van et al. [(1989) J. Biol. Chem. 264, 17494-17501]. These apparently homogeneous preparations showed only single 45Ca 2+ binding bands. On the basis of the peptide sequence, CBP2 was found to be highly homologous with the previously described protein ERp72. Similarly, CBP3 was identical to calreticulin and CBP1 had some homology to calmodulin. Contrary to the report of Van et al. (1989), we found that CBP2 had little thiol:protein disulfide oxidoreductase activity. Of the three purified preparations, only CBP2 exhibited apparent intrinsic protein kinase activity. This activity was found to be due to contamination of the CBP2 preparation by an extremely low concentration of tightly bound casein kinase 2 (CK2). In line with this observation, the phosphorylation was inhibited by heparin, removed by antibody to CK2, and stimulated by spermine. Furthermore, CBP2 was readily phosphorylated in vitro by added CK2 but only slowly phosphorylated by several other protein kinases. Thus, the persistence of CK2 in a highly purified preparation of CBP2 along with several other lines of evidence presented in this study might suggest that the protein CBP2 is a physiologically relevant substrate for CK2. Furthermore, these data suggest that CK2 might be localized in the lumen of the endoplasmic reticulum and that the phosphorylation of CBP2 in the lumen may play a role in the chaperone activity attributed to this protein.

Original languageEnglish (US)
Pages (from-to)8299-8306
Number of pages8
JournalBiochemistry
Volume35
Issue number25
DOIs
StatePublished - 1996

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    Chen, N., Davis, A. T., Canbulat, E. C., Liu, Y., Goueli, S., McKenzie, B. A., Eccleston, E. D., Ahmed, K., & Holtzman, J. L. (1996). Evidence that casein kinase 2 phosphorylates hepatic microsomal calcium- binding proteins 1 and 2 but not 3. Biochemistry, 35(25), 8299-8306. https://doi.org/10.1021/bi960296e