Please use this identifier to cite or link to this item: http://biore.bio.bg.ac.rs/handle/123456789/1072
Title: Biochemical characterization of soluble nucleotide pyrophosphatase/ phosphodiesterase activity in rat serum
Authors: Laketa, Danijela 
Bjelobaba, Ivana
Savic, Jasmina
Lavrnja, Irena
Stojiljkovic, Mirjana
Rakic, Ljubisav
Nedeljković, Nadežda 
Keywords: ATP analogs;NPP;Nucleotide pyrophosphatase/phosphodiesterase;Purinergic signaling;Rat serum
Issue Date: 1-Jun-2010
Journal: Molecular and Cellular Biochemistry
Abstract: 
Biochemical properties of nucleotide pyrophosphatase/phosphodiesterase (NPP) in rat serum have been described by assessing its nucleotide phosphodiesterase activity, using p-nitrophenyl-5′-thymidine monophosphate (p-Nph-5′-TMP) as a substrate. It was demonstrated that NPP activity shares some typical characteristics described for other soluble NPP, such as divalent cation dependence, strong alkaline pH optimum (pH 10.5), inhibition by glycosaminoglycans, and K m for p-Nph-5′-TMP hydrolysis of 61.8 ± 5.2 μM. In order to characterize the relation between phosphodiesterase and pyrophosphatase activities of NPP, we have analyzed the effects of different natural nucleotides and nucleotide analogs. ATP, ADP, and AMP competitively inhibited p-Nph-5′-TMP hydrolysis with K i values ranging 13-43 μM. Nucleotide analogs, α,β-metATP, BzATP, 2-MeSATP, and dialATP behaved as competitive inhibitors, whereas α,β-metADP induced mixed inhibition, with K i ranging from 2 to 20 μM. Chromatographic analysis revealed that α,β-metATP, BzATP, and 2-MeSATP were catalytically degraded in the serum, whereas dialATP and α,β-metADP resisted hydrolysis, implying that the former act as substrates and the latter as true competitive inhibitors of serum NPP activity. Since NPP activity is involved in generation, breakdown, and recycling of extracellular adenine nucleotides in the vascular compartment, the results suggest that both hydrolyzable and non-hydrolyzable nucleotide analogs could alter the amplitude and direction of ATP actions and could have potential therapeutic application. © 2010 Springer Science+Business Media, LLC.
URI: http://hdl.handle.net/123456789/1072
ISSN: 03008177
DOI: 10.1007/s11010-009-0373-1
Appears in Collections:Journal Article

Show full item record

SCOPUSTM   
Citations

11
checked on Sep 24, 2021

Page view(s)

5
checked on Feb 7, 2020

Google ScholarTM

Check

Altmetric

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.