Calcium-activated, phospholipid-dependent protein kinase in pancreatic acinar cells. 1985

M Noguchi, and H Adachi, and J D Gardner, and R T Jensen

In the present study we partially purified calcium-activated, phospholipid-dependent protein kinase (protein kinase C) from pancreatic acinar cells of the guinea pig using diethylaminoethylcellulose and Sephadex G-150 chromatography and characterized the dependence of the enzyme on calcium, phospholipids, diacylglycerol (diolein), and the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA). The enriched preparation of protein kinase C contained no cyclic nucleotide-dependent or calcium-dependent, calmodulin-dependent protein kinase activity. The values of Km for H1-histone and ATP were 0.74 +/- 0.22 and 13.1 +/- 3.2 microM, respectively. Pancreatic protein kinase C demonstrated an absolute requirement for calcium and phospholipid for its activation, and diolein or TPA increased the affinity of the enzyme for calcium by 10-fold. With phosphatidylserine the calcium concentration that caused a half-maximal activation (Ka) was 74 +/- 17 microM, whereas with phosphatidylserine and diolein or TPA the Ka for calcium was 7.9 +/- 1.6 or 6.8 +/- 1.3 microM, respectively. Adding phosphatidylethanolamine and phosphatidylserine decreased the Ka for calcium to 2.0 +/- 0.9 microM with diolein and to 0.7 +/- 0.4 microM with TPA. Activation of protein kinase C by TPA and diolein was identical with calcium concentrations greater than 1 microM, but at low calcium concentrations (less than 1 microM) in the presence of phospholipids, maximally effective concentrations of diolein caused only 55% of the activation seen with TPA. In addition to TPA, other phorbol esters such as phorbol dibutyrate and phorbol diacetate, but not phorbol itself, activated protein kinase C.(ABSTRACT TRUNCATED AT 250 WORDS)

UI MeSH Term Description Entries
D007515 Islets of Langerhans Irregular microscopic structures consisting of cords of endocrine cells that are scattered throughout the PANCREAS among the exocrine acini. Each islet is surrounded by connective tissue fibers and penetrated by a network of capillaries. There are four major cell types. The most abundant beta cells (50-80%) secrete INSULIN. Alpha cells (5-20%) secrete GLUCAGON. PP cells (10-35%) secrete PANCREATIC POLYPEPTIDE. Delta cells (~5%) secrete SOMATOSTATIN. Islands of Langerhans,Islet Cells,Nesidioblasts,Pancreas, Endocrine,Pancreatic Islets,Cell, Islet,Cells, Islet,Endocrine Pancreas,Islet Cell,Islet, Pancreatic,Islets, Pancreatic,Langerhans Islands,Langerhans Islets,Nesidioblast,Pancreatic Islet
D008297 Male Males
D010703 Phorbol Esters Tumor-promoting compounds obtained from CROTON OIL (Croton tiglium). Some of these are used in cell biological experiments as activators of protein kinase C. Phorbol Diester,Phorbol Ester,Phorbol Diesters,Diester, Phorbol,Diesters, Phorbol,Ester, Phorbol,Esters, Phorbol
D010743 Phospholipids Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system. Phosphatides,Phospholipid
D011493 Protein Kinase C An serine-threonine protein kinase that requires the presence of physiological concentrations of CALCIUM and membrane PHOSPHOLIPIDS. The additional presence of DIACYLGLYCEROLS markedly increases its sensitivity to both calcium and phospholipids. The sensitivity of the enzyme can also be increased by PHORBOL ESTERS and it is believed that protein kinase C is the receptor protein of tumor-promoting phorbol esters. Calcium Phospholipid-Dependent Protein Kinase,Calcium-Activated Phospholipid-Dependent Kinase,PKC Serine-Threonine Kinase,Phospholipid-Sensitive Calcium-Dependent Protein Kinase,Protein Kinase M,Calcium Activated Phospholipid Dependent Kinase,Calcium Phospholipid Dependent Protein Kinase,PKC Serine Threonine Kinase,Phospholipid Sensitive Calcium Dependent Protein Kinase,Phospholipid-Dependent Kinase, Calcium-Activated,Serine-Threonine Kinase, PKC
D011494 Protein Kinases A family of enzymes that catalyze the conversion of ATP and a protein to ADP and a phosphoprotein. Protein Kinase,Kinase, Protein,Kinases, Protein
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D002848 Chromatography, DEAE-Cellulose A type of ion exchange chromatography using diethylaminoethyl cellulose (DEAE-CELLULOSE) as a positively charged resin. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) DEAE-Cellulose Chromatography,Chromatography, DEAE Cellulose,DEAE Cellulose Chromatography
D002850 Chromatography, Gel Chromatography on non-ionic gels without regard to the mechanism of solute discrimination. Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D004075 Diglycerides Glycerides composed of two fatty acids esterified to the trihydric alcohol GLYCEROL. There are two possible forms that exist: 1,2-diacylglycerols and 1,3-diacylglycerols. Diacylglycerol,Diacylglycerols

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