Protein kinase C phosphorylates two of the four known syndecan cytoplasmic domains in vitro.
The transmembrane heparan sulfate proteoglycans of the syndecan family are implicated to participate in several cellular reactions which are dependent on protein kinase C. We have used an in vitro assay to assess whether any of the Peptides corresponding to the complete cytoplasmic domains of rat syndecans 1 through 4 were used as substrates for the enzyme. The syndecan-2 (fibroglycan) and syndecan-3 (N-syndecan) peptides were both found to be phosphorylated by protein kinase C with Kms of 15 +/- 3 microM and 85 +/- 25 microM, respectively, while the syndecan-1 and -4 peptides were not phosphorylated under the conditions used. The sites of in vitro phosphorylation for syndecans-2 and -3 were localized to ser-197 and ser-339, respectively. Thus, among 13 available sites (serines and threonines) in the four peptides, two were selectively modified by the enzyme. The specificity and the kinetics of the reactions indicate that the cytoplasmic domains of syndecan-2 and -3 are likely to be physiological substrates for protein kinase C.
Prasthofer
T
T
Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Ek
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Ekman
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Owens
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R
Hook
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Johansson
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S
D000595:Amino Acid Sequence; D000818:Animals; D003593:Cytoplasm / Q000378:metabolism; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D011509:Proteoglycans / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D053668:Syndecan-1; D053669:Syndecan-2; D053670:Syndecan-3; D053671:Syndecan-4; D053667:Syndecans
D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D011509:Proteoglycans; C508243:Sdc1 protein, rat; C508247:Sdc2 protein, rat; C508303:Sdc3 protein, rat; C508307:Sdc4 protein, rat; D053668:Syndecan-1; D053670:Syndecan-3; D053671:Syndecan-4; D053667:Syndecans; D053669:Syndecan-2; D011493:Protein Kinase C