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Staurosporine-inhibitable protein kinase activity associated with secretory granule membranes isolated from rat submandibular gland cells

 Su Ryeon Seo  ;  Yeun Ju Kim  ;  Jeong Taeg Seo  ;  Syng-Ill Lee  ;  Dong Min Shin  ;  Hiroshi Sugiya  ;  Seok Jun Moon 
 ARCHIVES OF ORAL BIOLOGY, Vol.48(8) : 553-558, 2003 
Journal Title
Issue Date
Animals ; Calcium/pharmacology ; Carbazoles/pharmacology ; Dose-Response Relationship, Drug ; Enzyme Inhibitors/pharmacology* ; Indole Alkaloids ; Membrane Proteins/metabolism ; Phosphorylation/drug effects ; Protein Kinase C/antagonists & inhibitors ; Protein Kinase C/metabolism ; Protein Kinase Inhibitors ; Protein Kinases/metabolism* ; Rats ; Rats, Sprague-Dawley ; Secretory Vesicles/enzymology* ; Secretory Vesicles/ultrastructure ; Staurosporine/pharmacology* ; Submandibular Gland/enzymology*
Secretory granule membrane ; Staurosporine ; K252a ; Submandibular acinar cells
Protein kinases, such as protein kinase C, have been shown to be associated with secretory granules and to regulate the event of exocytosis in various tissues including parotid salivary acinar cells. However, in submandibular acinar cells that play an important role in the secretion of proteins into the oral cavity, kinase activity on the granule membrane has not been explored. Therefore, in the present study, we isolated the secretory granules from rat submandibular acinar cells and investigated the localisation of protein kinases on the granule membrane. Initially, we isolated and purified secretory granules from rat submandibular acinar cells. Addition of [γ-] ATP to granule-membrane lysate phosphorylated the granule-membrane-associated 26, 32, 55 and 58 kDa proteins, suggesting the presence of endogenous kinase activity on the membrane. Moreover, the phosphorylation of 26 and 32 kDa proteins was inhibited by staurosporine and K252a, both non-specific protein kinase C inhibitors. However, the phosphorylation of 26 and 32 kDa proteins was not inhibited by other protein kinase C inhibitors, such as calphostin C, GF109203X and chelerythrine, indicating that protein kinase C was not responsible for the phosphorylation. In addition, H-89, ML-9, KN-62 and genistein did not appear to inhibit this phosphorylation, indicating that protein kinase A, myosin light chain kinase (MLCK), Ca2+/calmodulin-dependent protein kinase II (CAMKII) and tyrosine kinase were not involved in the phosphorylation of 26 and 32 kDa proteins. Moreover, Ca2+ had no effect on the kinase activity. Therefore, our results suggest that an unidentified, staurosporine-inhibitable protein kinase activity is associated with the secretory granule membrane of rat submandibular acinar cells.
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2. College of Dentistry (치과대학) > Dept. of Oral Biology (구강생물학교실) > 1. Journal Papers
Yonsei Authors
Moon, Seok Jun(문석준) ORCID logo https://orcid.org/0000-0001-7282-2888
Seo, Jeong Taeg(서정택) ORCID logo https://orcid.org/0000-0003-2697-0251
Shin, Dong Min(신동민) ORCID logo https://orcid.org/0000-0001-6042-0435
Lee, Syng Ill(이승일)
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