Identification of two distinct E-NTPDases in liver of goldfish (Carassius auratus L.)

Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
K AllevaPablo J Schwarzbaum

Abstract

We have recently reported the existence of ATPase activity capable of hydrolyzing extracellular ATP and localized at the external cell membrane of goldfish hepatocytes [Am. J. Physiol. (1998) 274 R1031]. In the present study, we investigated whether one or more enzymes of the ATP diphosphohydrolase family (called E-NTPDases) are responsible for the hydrolysis of extracellular ATP and other nucleotides. Using soluble extracts from goldfish liver, enzyme activity was detected in the presence of ATP (32.1+/-4.0 nmol Pi liberated mg protein(-1) min(-1)), ADP (20.7+/-3.3 nmol Pi liberated mg protein(-1) min(-1)) and UTP (20.7+/-1.2 nmol Pi liberated mg protein(-1) min(-1)). In line with the presence of this hydrolytic activity, liver samples separated by non-denaturing gel electrophoresis and subsequently exposed to either ATP, ADP or UTP yielded a single band with enzyme activity and similar electrophoretic mobility. Subsequent SDS-PAGE electrophoresis of the active bands resulted in the appearance of two protein bands with molecular masses of 70 and 64 kDa. Immunoblotting of soluble extracts and microsomes obtained from goldfish liver, using a monoclonal antibody against CD39 (a well-known E-NTPDase), detected a single 97-kDa prot...Continue Reading

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Citations

Mar 6, 2009·Bulletin of Mathematical Biology·Osvaldo CharaPablo J Schwarzbaum
Aug 6, 2003·Life Sciences·Eduardo Pacheco RicoCarla Denise Bonan
Feb 19, 2009·Acta Physiologica·G Burnstock, A Verkhratsky
Mar 23, 2010·Journal of Theoretical Biology·Osvaldo CharaPablo J Schwarzbaum
Nov 20, 2009·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Denis Broock RosembergMaurício Reis Bogo
Feb 4, 2011·Journal of Experimental Zoology. Part A, Ecological Genetics and Physiology·Osvaldo CharaPablo J Schwarzbaum
Dec 8, 2004·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Vânia Lúcia VieiraMaria Rosa Chitolina Schetinger
Oct 24, 2002·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Jonathan A DranoffJean Sévigny
Apr 13, 2007·Physiological Reviews·Geoffrey Burnstock
May 4, 2011·Journal of Enzyme Inhibition and Medicinal Chemistry·Daniela PochmannLisiane De Oliveira Porciúncula

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