Purification, characterization and cellular localization of 5′-nucleotidase from <i>Torpedo</i> electric organ

Grondal, Ernst J.M.; Zimmermann, Herbert

doi:10.1042/bj2450805

Cited by 49 publications

(47 citation statements)

References 33 publications

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“…The activities were measured as described under "Experimental Procedures." appears that 1 mM EDTA provokes the complete inhibition of the two enzymes; this result indicates that these 5Ј-nucleotidases use Mg 2ϩ as a cofactor, as do the homologous enzymes described previously (39,40). The activity was restored by addition of 5 mM Mg 2ϩ (not shown).…”

Section: Analyses Of 5ј-nucleotidase Activity In the Different Body Psupporting

confidence: 73%

Catabolism of Pyrimidine Nucleotides in the Deep-sea Tube WormRiftia pachyptila

Pastra-Landis

Gaill

Hervé³

2002

Journal of Biological Chemistry

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The present study describes the distribution and properties of enzymes of the catabolic pathway of pyrimidine nucleotides in Riftia pachyptila, a tubeworm living around deep-sea hydrothermal vents and known to be involved in a highly specialized symbiotic association with a bacterium. The catabolic enzymes, 5-nucleotidase, uridine phosphorylase, and uracil reductase, are present in all tissues of the worm, whereas none of these enzymatic activities were found in the symbiotic bacteria. The 5-nucleotidase activity was particularly high in the trophosome, the symbiont-harboring tissue. These results suggest that the production of nucleosides in the trophosome may represent an alternative source of carbon and nitrogen for R. pachyptila, because these nucleosides can be delivered to other parts of the worm. This process would complement the source of carbon and nitrogen from organic metabolites provided by the bacterial assimilatory pathways. The localization of the enzymes participating in catabolism, 5-nucleotidase and uridine phosphorylase, and of the enzymes involved in the biosynthesis of pyrimidine nucleotides, aspartate transcarbamylase and dihydroorotase, shows a non-homogeneous distribution of these enzymes in the trophosome. The catabolic enzymes 5-nucleotidase and uridine phosphorylase activities increase from the center of the trophosome to its periphery. In contrast, the anabolic enzymes aspartate transcarbamylase and dihydroorotase activities decrease from the center toward the periphery of the trophosome. We propose a general scheme of anatomical and physiological organization of the metabolic pathways of the pyrimidine nucleotides in R. pachyptila and its bacterial endosymbiont.

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Section: Analyses Of 5ј-nucleotidase Activity In the Different Body Psupporting

confidence: 73%

Catabolism of Pyrimidine Nucleotides in the Deep-sea Tube WormRiftia pachyptila

Pastra-Landis

Gaill

Hervé³

2002

Journal of Biological Chemistry

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“…At equimolar substrate concentrations (0.5 mM; K,(AMP) for 5'-nucleotidase = 38 pM; Grondal and Zimmermann, 1987) the rate of hydrolysis of UDP-glucose was 8% of that observed for AMP (100% = 4.83 _+ 0.17 pmol Pi.min.mg protein-'; mean _+ SEM; IZ = 6).…”

Section: Activity Of Udp-glucose Hydrolase Associated With Electric Rmentioning

confidence: 90%

“…The recently described primary structures of 5'-nucleotidase from rat liver and human placenta (Misumi et al, 1990a,b) reveal similarities with the procaroytic enzymes. Here we report the primary structure deduced from cDNA of ecto-5'-nucleotidase from the electromotor system of electric ray, whose biochemical properties and tissue distribution have previously been described (Grondal and Zimmermann, 1987;. The electric ray enzyme bears a close similarity to the mammalian enzymes, but at the same time reveals surprising clusters of identity with procaryotic enzymes.…”

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confidence: 88%

5′‐Nucleotidase from the electric ray electric lobe

Volknandt

Vogel

Pevsner

et al. 1991

European Journal of Biochemistry

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A cDNA encoding a 5'-nucleotidase was identified by screening a lgtlO cDNA library from the electric lobe of Discopyge ommata using a cDNA probe containing the complete open reading frame coding for the rat liver enzyme. Nucleotide sequence analysis defines an open reading frame of 577 amino acids, corresponding to a calculated molecular mass of 63833 Da. The N-terminus of the mature protein, as determined by direct protein sequencing, is preceded by 29 amino acid residues comprising a signal peptide. The C-terminus contains a stretch of hydrophobic amino acids, considered to be cleaved on post-translational modification and exchanged for glycosylphosphatidylinositol as a membrane anchor. The predicted protein contains four potential N-linked glycosylation sites. Electric ray 5'-nucleotidase shares 61 % amino acid identity with the enzymes from rat liver and human placenta, and about 23 YO with bacterial proteins possessing 5'-nucleotidase activity and also additional enzyme activities like UDP-glucose hydrolase. Polyclonal antibodies raised against 5'-nucleotidase from mammalian sources or the electric ray electric organ reveal mutual crossreactivity. Interestingly, there are 5 -7 domains highly conserved in procaryotes and vertebrates in enzymes exhibiting 5'-nucleotidase, 3'-nucleotidase or phosphodiesterase activity. 5'-nucleotidase isolated from Torpedo electric organ hydrolyzes UDP-glucose at 8 % of the rate of AMP hydrolysis. The possible phylogenetic origin of vertebrate 5'-nucleotidase from multifunctional nucleotide hydrolases is discussed.

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“…Importantly, ATP and ADP are competitive inhibitors of vertebrate eN with K i values in the low micromolar range. These nucleotides apparently still bind to the catalytic site but without being hydrolyzed [209,210]. Cellular release of ATP/ADP would thus result in feed-forward inhibition of eN and delay of extracellular adenosine formation until their extracellular levels have been reduced to low micromolar levels by other ecto-nucleotidases [211].…”

Section: Ecto-5′-nucleotidasementioning

confidence: 99%

Cellular function and molecular structure of ecto-nucleotidases

Zimmermann

Zebisch

Sträter

2012

Purinergic Signalling

890

922

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Ecto-nucleotidases play a pivotal role in purinergic signal transmission. They hydrolyze extracellular nucleotides and thus can control their availability at purinergic P2 receptors. They generate extracellular nucleosides for cellular reuptake and salvage via nucleoside transporters of the plasma membrane. The extracellular adenosine formed acts as an agonist of purinergic P1 receptors. They also can produce and hydrolyze extracellular inorganic pyrophosphate that is of major relevance in the control of bone mineralization. This review discusses and compares four major groups of ectonucleotidases: the ecto-nucleoside triphosphate diphosphohydrolases, ecto-5′-nucleotidase, ecto-nucleotide pyrophosphatase/phosphodiesterases, and alkaline phosphatases. Only recently and based on crystal structures, detailed information regarding the spatial structures and catalytic mechanisms has become available for members of these four ecto-nucleotidase families. This permits detailed predictions of their catalytic mechanisms and a comparison between the individual enzyme groups. The review focuses on the principal biochemical, cell biological, catalytic, and structural properties of the enzymes and provides brief reference to tissue distribution, and physiological and pathophysiological functions.

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Purification, characterization and cellular localization of 5′-nucleotidase from Torpedo electric organ

Cited by 49 publications

References 33 publications

Catabolism of Pyrimidine Nucleotides in the Deep-sea Tube WormRiftia pachyptila

Catabolism of Pyrimidine Nucleotides in the Deep-sea Tube WormRiftia pachyptila

5′‐Nucleotidase from the electric ray electric lobe

Cellular function and molecular structure of ecto-nucleotidases

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