Summary
Endogenous calcium-activated, phospholipid-dependent protein kinase phosphorylates pig epidermal protein. Pig epidermis was homogenized and centrifuged at 30,000xg for 30 min. The supernatant was incubated with or without calcium and phospholipid. A 97 kD soluble protein from pig epidermis was phosphorylated in the presence of calcium and phosphatidylserine. The phosphorylation reaction occurred immediately. With the use of two-dimensional polyacrylamide gel electrophoresis, it was shown that the 97 kD fragment was a basic protein, and that several small proteins were also phosphorylated. The characterization of these proteins is yet to be undertaken.
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Castagna M, Takai Y, Kaibuchi K, Sano K, Kikkawa U, Nishizuka Y (1982) Direct activation of calcium-activated phospholipid-dependent protein kinase by tumor-promoting phorbol esters. J Biol Chem 257:7847–7851
Endo T, Naka M, Hidaka H (1982) Ca2+-phospholipid dependent phosphorylation of smooth muscle myosin. Biochem Biophys Res Commun 105:942–948
Feuerstein N, Cooper HL (1984) Rapid phosphorylation of specific proteins induced by phorbol ester in HL-60 cells. J Biol Chem 259:2782–2788
Iwasa Y, Hosey MM (1984) Phosphorylation of cardiac sarcolemma proteins by the calcium-activated phospholipid-dependent protein kinase. J Biol Chem 259:534–540
Katada T, Gilman AG, Watanabe Y, Bauer S, Jakobs KH (1985) Protein kinase C phosphorylates the inhibitory guanine-nucleotide-binding regulatory component and apparently suppress its function in hormonal inhibition of adenylate cyclase. Eur J Biol 151:431–437
Katoh N, Kuo JF (1982) Subcellular distribution of phospholipid-sensitive calcium-dependent protein kinase in guinea-pig heart, spleen, and cerebral cortex, and inhibition of the enzyme by Triton X-100. Biochem Biophys Res Commun 106:590–595
Katoh N, Wise BC, Kuo JF (1983) Phosphorylation of cardiac troponin inhibitory subunit (troponin I) and tropomyosin-binding subunit (troponin T) by cardiac phospholipid-sensitive Ca2+-dependent protein kinase. Biochem J 209:189–195
Kikkawa U, Takai Y, Minakuchi R, Inohara S, Nishizuka Y (1982) Calcium-activated, phospholipid-dependent protein kinase from rat brain. Subcellular distribution, purification and properties. J Biol Chem 257:13341–13348
Kimura K, Katoh N, Sakurada K, Kubo S (1984) Phospholipid-sensitive Ca2+-dependent protein kinase system in testis: Localization and endogenous substrates. Endocrinology 115:2391–2399
Koizumi H, Adachi K, Tsutsui M, Itami S, Katayama H, Halprin KM (1984) Calcium-activated, phospholipid-dependent protein kinase in pig epidermis. J Invest Dermatol 83:261–264
Kraft AS, Anderson WB (1983) Characterization of cytosolic calcium-activated phospholipid-dependent protein kinase activity in embryonal carcinoma cells. Effect of retinoic acid-induced differentiation of F-9 cells to parietal endoderm. J Biol Chem 258:9178–9183
Kuo JF, Andersson RGG, Wise BC, Mackerlova L, Salommonsson I, Brackett NL, Katoh N, Shoji M, Wrenn RW (1980) Calcium-dependent protein kinase: wide-spread occurrence in various tissues and phyla of the animal kingdom and comparison of effects of phospholipid, calmodulin and trifluoperazine. Proc Natl Acad Sci USA 77:7039–7043
Laemmli UK (1970) Cleavage of structural protein during assembly of the head of bacteriophage T4. Nature 227:680–685
LePeuch CJ, Ballester R, Rosen OM (1983) Purified rat brain calcium- and phospholipid-dependent protein kinase phosphorylates ribosomal protein S6. Proc Natl Acad Sci USA 80:6858–6862
Limas CJ, Limas C (1985) Phorbol ester- and diacylglycerolmediated desensitization of cardiac β-adrenergic receptors. Circ Res 57:443–449
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin phenol reagent. J Biol Chem 193:265–275
Nishikawa M, Hidaka H, Adelson RS (1983) Phosphorylation of smooth muscle heavy meromyosin by calcium-activated, phospholipid-dependent protein kinase. J Biol Chem 258:14069–14072
Nishizuka Y (1984) Turnover of inositol phospholipids and signal transduction. Science 225:1365–1370
O'Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021
Papanicolaou P, Humble E, Engstrom L (1982) Phosphorylation of human fibrinogen in vitro with calcium-activated phospholipid-dependent protein kinase and (32P) ATP. FEBS Lett 143:199–204
Sakurada K, Miyazaki T, Kimura K, Yamabe M, Katabami F, Koyama M, Uehara Y, Katoh N (1985) Translocation of phospholipid-sensitive Ca2+-dependent protein kinase and its substrate, Mr 38,000 protein, in chronic myelocytic and acute myelocytic leukemias. Cancer Res 45:903–908
Turner RS, Chou CHJ, Kibler RF, Kuo JF (1982) Basic protein in brain myelin is phosphorylated by endogenous phospholipid-sensitive Ca2+-dependent protein kinase. J Neurochem 39:1397–1404
Werth DK, Niedel JE, Plastan I (1983) Vinculin, a cytoskeletal substrate of protein kinase C. J Biol Chem 258:11423–11426
Wu WCS, Walaas SI, Nairn AC, Greencard P (1982) Calcium/phospholipid regulates phosphorylation of a Mr “87 k” substrate protein in brain synaptosome. Proc Natl Acad Sci USA 79:5249–5253
Yoshikawa K, Takeda J, Nemoto O, Ito T, Halprin KM, Adachi K (1983) Phosphorylation of pig epidermal soluble protein by endogenous cAMP-dependent protein kinase. J Invest Dermatol 80:108–111
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Koizumi, H., Aoyagi, T. & Ohkawara, A. Phosphorylation of soluble pig epidermal proteins by endogenous calcium-activated, phospholipid-dependent protein kinase. Arch Dermatol Res 279, 95–99 (1986). https://doi.org/10.1007/BF00417529
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DOI: https://doi.org/10.1007/BF00417529