Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/2931
Title: Purification, characterization, and localization of an ATP diphosphohydrolase in porcine kidney
Authors: LEMMENS, Raf 
KUPERS, Luc 
Sevigny, J
Beaudoin, A.R.
Grondin, G
Kittel, A
Waelkens, E
VANDUFFEL, Luc 
Issue Date: 2000
Publisher: AMER PHYSIOLOGICAL SOC
Source: AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 278(6). p. F978-F988
Abstract: Membranes of pig kidney cortex tissue were solubilized in the presence of Triton X-100. Partial purification of ATP diphosphohydrolase (ATPDase) was achieved by successive chromatography on concanavalin A-Sepharose, Q-Sepharose Fast Flow, and 5'-AMP-Sepharose 4B. Monoclonal antibodies against ATPDase were generated. Further purification of the ATPDase was obtained by immunoaffinity chromatography with these monoclonal antibodies. NH2-terminal amino acid sequencing of the 78-kDa protein showed a sequence very homologous to mammalian CD39. The protein is highly glycosylated, with a nominal molecular mass of similar to 57 kDa. The purified enzyme hydrolyzed di- and triphosphates of adenosine, guanosine, cytidine, uridine, inosine, and thymidine, but AMP and diadenosine polyphosphates could not serve as substrates. All enzyme activities were dependent on divalent cations and were partially inhibited by 10 mM sodium azide. The distribution of the enzyme in pig kidney cortex was examined immunohistochemically. The enzyme was found to be present in blood vessel walls of glomerular and peritubular capillaries.
Notes: Limburgs Univ Ctr, Dept MBW, B-3590 Diepenbeek, Belgium. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Med, Boston, MA 02215 USA. Univ Sherbrooke, Dept Biol, Sherbrooke, PQ J1K 2R1, Canada. Hungarian Acad Sci, Inst Expt Med, H-1450 Budapest, Hungary. Katholieke Univ Leuven, B-3000 Louvain, Belgium.Vanduffel, L, Limburgs Univ Ctr, Dept MBW, Univ Campus,Bldg D, B-3590 Diepenbeek, Belgium.
Keywords: CD39; apyrase; nucleoside triphosphate diphosphohydrolase; Triton X-100
Document URI: http://hdl.handle.net/1942/2931
ISI #: 000087573900014
Category: A1
Type: Journal Contribution
Validations: ecoom 2001
Appears in Collections:Research publications

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