© 1994 Oxford University Press
research-article |
12-Deoxyphorbol-13-O-phenylacetate-20-acetate is not protein kinase C-ß isozyme-selective in vivo
Protein Phosphorylation Laboratory, Imperial Cancer Research Fund PO Box 123, Lincoln's Inn Fields, London WC2A 3PX
1Department of Pharmacognosy, The School of Pharmacy, University of London 29-39 Brunswick Square, London WC1N 1AX, UK
2Division of Endocrinology, Department of Medicine, East Carolina University School of Medicine Greenville, NC 27858–4354, USA
3To whom correspondence should be addressed
The phorbol ester, 12-deoxyphorbol-13-O-phenylacetate-20-acetate (DOPPA) has been shown to activate specifically the protein kinase C (PKQ-ß1 isozyme in vitro (1). We have investigated the potential of DOPPA as a PKC-ß
isozyme-specific agonist hi intact cells, employing U937 cells, which express ß, 
and 
PKC and in Swiss 3T3 cells which lack PKC-ß but express 
, 
, and PKC. Immunoblot analysis with isozyme-specific antibodies indicated that DOPPA can mediate the subcellular redistribution and down-modulation of all endogenous PKC isozymes (except PKC-) in both U937 and Swiss 3T3 cells. Prolonged treatment (>6 h) of cultures in down-modulation studies is complicated by the metabolism of DOPPA to 12-deoxyphorbol-13-phenylacetate (DOPP), a compound which activates all PKC isozymes tested in vitro (Ryves,WJ. et al. (1991) FEBS Lett., 288, 5–9). Nevertheless, because DOPPA induced rapid and dose-dependent phosphorylation of p80 in cells which do not express PKC-ß, p80 phosphorylation in Swiss 3T3 cells indicates that DOPPA can activate a non-ß PKC in vivo. The data suggest that DOPPA cannot be used as a PKC-ß-selective agonist in intact cell studies.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. R. Broughman, L. Sun, S. Umar, J. Scott, J. H. Sellin, and A. P. Morris Chronic PKC-beta activation in HT-29 Cl.19a colonocytes prevents cAMP-mediated ion secretion by inhibiting apical membrane current generation Am J Physiol Gastrointest Liver Physiol, August 1, 2006; 291(2): G318 - G330. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. R. Broughman, L. Sun, S. Umar, J. H. Sellin, and A. P. Morris Chronic PKC-beta2 activation in HT-29 Cl.19a colonocytes prevents cAMP-mediated ion secretion by inhibiting apical membrane CFTR targeting Am J Physiol Gastrointest Liver Physiol, August 1, 2006; 291(2): G331 - G344. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Husoy, V. Cruciani, T. Sanner, and S.-O. Mikalsen Phosphorylation of connexin43 and inhibition of gap junctional communication in 12-O-tetradecanoylphorbol-13-acetate-exposed R6 fibroblasts: minor role of protein kinase C{beta}I and {micro} Carcinogenesis, February 1, 2001; 22(2): 221 - 231. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Tian, J. A C Philp, and M. J Shipston Glucocorticoid block of protein kinase C signalling in mouse pituitary corticotroph AtT20 D16:16 cells J. Physiol., May 1, 1999; 516(3): 757 - 768. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. B. King, A. Norvell, and J. G. Monroe Antigen Receptor-Induced Signal Transduction Imbalances Associated with the Negative Selection of Immature B Cells J. Immunol., March 1, 1999; 162(5): 2655 - 2662. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. C. Das, X.-l. Guo, and C. W. White Protein Kinase Cdelta -dependent Induction of Manganese Superoxide Dismutase Gene Expression by Microtubule-active Anticancer Drugs J. Biol. Chem., December 18, 1998; 273(51): 34639 - 34645. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. C. Mason, H. Yarwood, K. Sugars, and D. O. Haskard Human umbilical vein and dermal microvascular endothelial cells show heterogeneity in response to PKC activation Am J Physiol Cell Physiol, October 1, 1997; 273(4): C1233 - C1240. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Kiley and P. Parker Differential localization of protein kinase C isozymes in U937 cells: evidence for distinct isozyme functions during monocyte differentiation J. Cell Sci., January 3, 1995; 108(3): 1003 - 1016. [Abstract] [PDF]
|
|