title: Evidence that casein kinase 2 phosphorylates hepatic microsomal calcium-binding proteins 1 and 2 but not 3.
abstract: We have extensively purified three of the hepatic microsomal intralumenal Ca2+-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 45Ca2+ 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.
authors.affiliation: Medical and Research Services, Veterans Affairs Medical Center, Minneapolis, Minnesota 55417, USA.
authors.forename: N Q
authors.initials: NQ
authors.lastname: Chen
authors.forename: A T
authors.initials: AT
authors.lastname: Davis
authors.forename: E C
authors.initials: EC
authors.lastname: Canbulat
authors.forename: Y X
authors.initials: YX
authors.lastname: Liu
authors.forename: S
authors.initials: S
authors.lastname: Goueli
authors.forename: B A
authors.initials: BA
authors.lastname: McKenzie
authors.forename: E D
authors.initials: ED
authors.lastname: Eccleston
authors.forename: K
authors.initials: K
authors.lastname: Ahmed
authors.forename: J L
authors.initials: JL
authors.lastname: Holtzman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037282:Calreticulin; D047390:Casein Kinase II; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012261:Ribonucleoproteins / Q000378:metabolism*; D017421:Sequence Analysis; D013379:Substrate Specificity
chemical_list: D002135:Calcium-Binding Proteins; D037282:Calreticulin; D008562:Membrane Glycoproteins; D012261:Ribonucleoproteins; C100371:calcium-binding protein-1 (liver); C062477:endoplasmic reticulum glycoprotein p72; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases