AJP - Cell Physiology, Vol 267, Issue 4 C1073-C1079, Copyright © 1994 by American Physiological Society

ARTICLES

Decreased protein phosphorylation induced by anoxia in proximal renal tubules

C. E. Kobryn and L. J. Mandel
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710.

Anoxia-induced depletion of cellular ATP may affect the degree of protein phosphorylation due to kinase inhibition. In this study, protein phosphorylation was measured in rabbit kidney proximal tubules under normoxic or anoxic conditions in a medium containing 32P. During the first 20 min of normoxia, phosphate incorporation was linear, averaging 17 +/- 5 pmol.mg protein-1.min-1 and was 70% inhibited by the protein kinase C inhibitor chelerythrine chloride. Phosphorylation measurements initiated simultaneously with anoxic conditions (95% N2-5% CO2) significantly reduced the initial rate to 58% of control, saturating after 15 min, and reaching 28 +/- 5% of the normoxic value after 60 min of incubation. The phosphatase inhibitor calyculin A did not affect the initial rate of phosphate incorporation by anoxic tubules but increased phosphate incorporation at 60 min to 43 +/- 4% of normoxia. Addition of 32P after 15 min of anoxia abolished phosphate incorporation, demonstrating that kinase activity was completely inhibited. Cellular phosphate uptake was measured and found not to be rate limiting for phosphorylation. Chelerythrine chloride increased lactate dehydrogenase (LDH) release during normoxia, and calyculin A decreased anoxia-induced LDH release, suggesting that protein phosphorylation events may control plasma membrane permeability.

This article has been cited by other articles:

				S. Gopalakrishnan, M. A. Hallett, S. J. Atkinson, and J. A. Marrs aPKC-PAR complex dysfunction and tight junction disassembly in renal epithelial cells during ATP depletion Am J Physiol Cell Physiol, March 1, 2007; 292(3): C1094 - C1102. [Abstract] [Full Text] [PDF]

				J. V. Bonventre and J. M. Weinberg Recent Advances in the Pathophysiology of Ischemic Acute Renal Failure J. Am. Soc. Nephrol., August 1, 2003; 14(8): 2199 - 2210. [Full Text] [PDF]

				S. Gopalakrishnan, K. W. Dunn, and J. A. Marrs Rac1, but not RhoA, signaling protects epithelial adherens junction assembly during ATP depletion Am J Physiol Cell Physiol, July 1, 2002; 283(1): C261 - C272. [Abstract] [Full Text] [PDF]

				Z.-H. Zhou and J. K. Bubien Nongenomic regulation of ENaC by aldosterone Am J Physiol Cell Physiol, October 1, 2001; 281(4): C1118 - C1130. [Abstract] [Full Text] [PDF]

				J. M. Weinberg, M. A. Venkatachalam, N. F. Roeser, R. A. Senter, and I. Nissim Energetic Determinants of Tyrosine Phosphorylation of Focal Adhesion Proteins during Hypoxia/Reoxygenation of Kidney Proximal Tubules Am. J. Pathol., June 1, 2001; 158(6): 2153 - 2164. [Abstract] [Full Text] [PDF]

				J. H. SCHWARTZ, T. SHIH, S. A. MENZA, and W. LIEBERTHAL ATP Depletion Increases Tyrosine Phosphorylation of {beta}-Catenin and Plakoglobin in Renal Tubular Cells J. Am. Soc. Nephrol., November 1, 1999; 10(11): 2297 - 2305. [Abstract] [Full Text]

				N. Raman and S. J. Atkinson Rho controls actin cytoskeletal assembly in renal epithelial cells during ATP depletion and recovery Am J Physiol Cell Physiol, June 1, 1999; 276(6): C1312 - C1324. [Abstract] [Full Text] [PDF]

				S. Gopalakrishnan, N. Raman, S. J. Atkinson, and J. A. Marrs Rho GTPase signaling regulates tight junction assembly and protects tight junctions during ATP depletion Am J Physiol Cell Physiol, September 1, 1998; 275(3): C798 - C809. [Abstract] [Full Text] [PDF]

				A. M. Sheridan, A. Sapirstein, N. Lemieux, B. D. Martin, D. K. Kim, and J. V. Bonventre Nuclear Translocation of Cytosolic Phospholipase A2 Is Induced by ATP Depletion J. Biol. Chem., August 3, 2001; 276(32): 29899 - 29905. [Abstract] [Full Text] [PDF]