High-Performance Liquid Chromatographic Assay of the Diadenosine Polyphosphates in Human Platelets

Jankowski, Joachim; Potthoff, Wilhelm; Giet, Markus van der; Tepel, Martín; Zidek, Walter; Schlüter, Hartmut

doi:10.1006/abio.1999.3097

Cited by 28 publications

(20 citation statements)

References 23 publications

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“…The intracellular concentration of Ap 4 A in unstressed cells is typically 0.1-1.0 M (46), whereas PP-InsP 5 may be in the low micromolar range (47). There are no measurements of cytoplasmic Ap 6 A in mammalian cells; an estimate of 32 nM in platelets is an intracellular average because the Ap 6 A is concentrated in the dense granules (48). Indeed, it is likely to be even lower than the sole measured value for Ap 5 A of 4 nM in Schizosaccharomyces pombe (52).…”

Section: Discussionmentioning

confidence: 99%

Nudix Hydrolases That Degrade Dinucleoside and Diphosphoinositol Polyphosphates Also Have 5-Phosphoribosyl 1-Pyrophosphate (PRPP) Pyrophosphatase Activity That Generates the Glycolytic Activator Ribose 1,5-Bisphosphate

Fisher

Safrany

Strike

et al. 2002

Journal of Biological Chemistry

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A total of 17 Nudix hydrolases were tested for their ability to hydrolyze 5-phosphoribosyl 1-pyrophosphate (PRPP). All 11 enzymes that were active toward dinucleoside polyphosphates with 4 or more phosphate groups as substrates were also able to hydrolyze PRPP, whereas the 6 that could not and that have coenzyme A, NDP-sugars, or pyridine nucleotides as preferred substrates did not degrade PRPP. The products of hydrolysis were ribose 1,5-bisphosphate and P i . Active PRPP pyrophosphatases included the diphosphoinositol polyphosphate phosphohydrolase (DIPP) subfamily of Nudix hydrolases, which also degrade the non-nucleotide diphosphoinositol polyphosphates. K m and k cat values for PRPP hydrolysis for the Deinococcus radiodurans DR2356 (di)nucleoside polyphosphate hydrolase, the human diadenosine tetraphosphate hydrolase, and human DIPP-1 (diadenosine hexaphosphate and diphosphoinositol polyphosphate hydrolase) were 1 mM and 1.5 s ؊1 , 0.13 mM and 0.057 s ؊1 , and 0.38 mM and 1.0 s ؊1 , respectively. Active site mutants of the Caenorhabditis elegans diadenosine tetraphosphate hydrolase had no activity, confirming that the same active site is responsible for nucleotide and PRPP hydrolysis. Comparison of the specificity constants for nucleotide, diphosphoinositol polyphosphate, and PRPP hydrolysis suggests that PRPP is a significant substrate for the D. radiodurans DR2356 enzyme and for the DIPP subfamily. In the latter case, generation of the glycolytic activator ribose 1,5-bisphosphate may be a new function for these enzymes.

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Section: Discussionmentioning

confidence: 99%

Nudix Hydrolases That Degrade Dinucleoside and Diphosphoinositol Polyphosphates Also Have 5-Phosphoribosyl 1-Pyrophosphate (PRPP) Pyrophosphatase Activity That Generates the Glycolytic Activator Ribose 1,5-Bisphosphate

Fisher

Safrany

Strike

et al. 2002

Journal of Biological Chemistry

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“…In the platelets contained in 1 L of whole blood, the following amounts of diadenosine polyphosphates were found 44 : Ap 3 A, 192.5Ϯ14.7 nmol; Ap 4 A, 223.8Ϯ16.8 nmol; Ap 5 A, 100.2Ϯ7.9 nmol; Ap 6 A, 32.0Ϯ1.9 nmol (mean Ϯ SEM). Conceivably, even if 100% of the platelet diadenosine polyphosphates are assumed to be released and hence to be distributed within the pertinent volume, the resulting plasma concentrations would be far less than those reported here.…”

Section: Jankowski Et Al Diadenosine Polyphosphates Adrenal Glands mentioning

confidence: 97%

“…Although this question cannot be solved on the basis of the present data, findings reported in the literature may give an answer: concerning platelets, several studies revealed that the diadenosine polyphosphate contents decreased with an increasing number of phosphate groups. 42,44 Because the intraplatelet diadenosine polyphosphates are not accessible by the extracellular degrading enzymes, it appears more likely that the synthetic pathway is less effective with increasing number of phosphate moieties to be incorporated.…”

Section: Jankowski Et Al Diadenosine Polyphosphates Adrenal Glands mentioning

confidence: 99%

Identification and Quantification of Diadenosine Polyphosphate Concentrations in Human Plasma

Jankowski

Laufer

et al. 2003

ATVB

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Objective-Diadenosine polyphosphates have been demonstrated to be involved in the control of vascular tone as well as the growth of vascular smooth muscle cells and hence, possibly, in atherogenesis. In this study we investigated the question of whether diadenosine polyphosphates are present in human plasma and whether a potential source can be identified that may release diadenosine polyphosphates into the circulation. Methods and Results-Plasma diadenosine polyphosphates (Ap n A with nϭ3 to 6) were purified to homogeneity by affinity-, anion exchange-, and reversed phase-chromatography from deproteinized human plasma. Analysis of the homogeneous fractions with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) revealed molecular masses ([MϩH] ϩ ) of 757, 837, 917, and 997 d. Comparison of the postsource decay matrix-assisted laser desorption/ionization mass spectrometry mass spectra of these fractions with those of authentic diadenosine polyphosphates revealed that these isolated substances were identical to Ap 3 A, Ap 4 A, Ap 5 A, and Ap 6 A. Enzymatic analysis showed an interconnection of the phosphate groups with the adenosines in the 5Ј-positions of the ribose moieties. The mean total plasma diadenosine polyphosphate concentrations (mol/L; mean Ϯ SEM) in cubital veins of normotensive subjects amounted to 0.89Ϯ0.59 for Ap 3 A, 0.72Ϯ0.72 for Ap 4 A, 0.33Ϯ0.24 for Ap 5 A, and 0.18Ϯ0.18 for Ap 6 A. Cubital venous plasma diadenosine polyphosphate concentrations from normotensives did not differ significantly from those in the hypertensive patients studied. There was no significant difference between arterial and venous diadenosine polyphosphate plasma concentrations in 5 hemodialysis patients, making a significant degradation by capillary endothelial cells unlikely. Free plasma diadenosine polyphosphate concentrations are considerably lower than total plasma concentrations because approximately 95% of the plasma diadenosine polyphosphates were bound to proteins. There were no significant differences in the diadenosine polyphosphate plasma concentrations depending on the method of blood sampling and anticoagulation, suggesting that platelet aggregation does not artificially contribute to plasma diadenosine polyphosphate levels in significant amounts.The Ap n A (with nϭ3 to 6) total plasma concentrations in adrenal veins were significantly higher than the plasma concentrations in both infrarenal and suprarenal vena cava

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“…Another example is the use of a Cu 2+ -affinity column with MS for purifying and examining P2 protein from myelin cells [205]. In addition, boronate columns have been used in off-line affinity extraction with RP chromatography for the analysis of urinary nucleosides in intestinal cancer patients [206], and for the concentration of diadenosine pentaphosphate and diadenosine hexaphosphate in human platelets prior to quantitation by HPLC and MS [207]. Use of nickel as an affinity ligand for the purification of recombinant human prolactin has been reported as well [208], and immobilized iron (III) has been used to isolate phosphopeptides from the bovine adrenal medulla gland [209].…”

Section: Biointeraction Studiesmentioning

confidence: 99%

Applications of silica supports in affinity chromatography

Schiel

Mallik

Soman

et al. 2006

J of Separation Science

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The combined use of silica-based chromatographic supports with immobilized affinity ligands can be used in many preparative and analytical applications. One example is the use of silica-based affinity columns in HPLC, giving rise to a method known as high-performance affinity chromatography (HPAC). This review discusses the role that silica has played in the development of affinity chromatography and HPAC and the applications of silica in these methods. This includes a discussion of the types of ligands that have been employed with silica and the methods by which these ligands have been immobilized. Various formats have also been presented for the use of silica in affinity chromatographic methods, including assays involving direct or indirect analyte detection, on-line or off-line affinity extraction, and chiral separations. The use of silica-based affinity columns in studies of biological systems based on zonal elution and frontal analysis methods will also be considered.

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High-Performance Liquid Chromatographic Assay of the Diadenosine Polyphosphates in Human Platelets

Cited by 28 publications

References 23 publications

Nudix Hydrolases That Degrade Dinucleoside and Diphosphoinositol Polyphosphates Also Have 5-Phosphoribosyl 1-Pyrophosphate (PRPP) Pyrophosphatase Activity That Generates the Glycolytic Activator Ribose 1,5-Bisphosphate

Nudix Hydrolases That Degrade Dinucleoside and Diphosphoinositol Polyphosphates Also Have 5-Phosphoribosyl 1-Pyrophosphate (PRPP) Pyrophosphatase Activity That Generates the Glycolytic Activator Ribose 1,5-Bisphosphate

Identification and Quantification of Diadenosine Polyphosphate Concentrations in Human Plasma

Applications of silica supports in affinity chromatography

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