Identification and characterization of P(1), P(7)-Di(adenosine-5')-heptaphosphate from human platelets
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Identification and characterization of P(1), P(7)-Di(adenosine-5')-heptaphosphate from human platelets. / Jankowski, J; Tepel, M; van der Giet, M; Tente, I M; Henning, L; Junker, R; Zidek, W; Schlüter, H.
In: J BIOL CHEM, Vol. 274, No. 34, 20.08.1999, p. 23926-31.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Identification and characterization of P(1), P(7)-Di(adenosine-5')-heptaphosphate from human platelets
AU - Jankowski, J
AU - Tepel, M
AU - van der Giet, M
AU - Tente, I M
AU - Henning, L
AU - Junker, R
AU - Zidek, W
AU - Schlüter, H
PY - 1999/8/20
Y1 - 1999/8/20
N2 - Diadenosine pentaphosphate and diadenosine hexaphosphate have been isolated in human platelets and have been postulated to play an important role in the control of vascular tone. Here we describe the isolation and identification of diadenosine heptaphosphate from human platelets. Dinucleoside polyphosphates were concentrated by affinity chromatography from a nucleotide-containing fraction from deproteinated human platelets. Dinucleoside polyphosphates were purified by anion-exchange and reversed phase high performance liquid chromatography to homogeneity. Analysis of one of these fractions with matrix-assisted laser desorption/ionization mass spectrometry revealed a molecular mass of 1076.4 (1077.4 = [M + H](+)) Da. UV spectroscopic analysis of this fraction showed the spectrum of an adenosine derivative. Comparison of the postsource decay matrix-assisted laser desorption/ionization mass spectrum of the fraction minus that of diadenosine heptaphosphate (Ap(7)A) demonstrated that the isolated substance was identical to Ap(7)A. The identity of the retention times of the authentic and the isolated compound confirmed this result. Enzymatic analysis demonstrated an interconnection of the phosphate groups with the adenosines in the 5'-positions of the riboses. With thrombin-induced platelet aggregation, Ap(7)A is released from the platelets into the extracellular space. The vasoconstrictive action of Ap(7)A on the vasculature of the isolated perfused rat kidney Ap(7)A was slightly less than that of Ap(6)A. The threshold of the vasoconstrictive action of Ap(7)A was 10(-5) mol/liter. The vasoconstrictive effect was abolished by suramin and pyridoxal phosphate 6-azophenyl-2', 4'-disulfonic acid, suggesting an activation of P(2x) receptors. Furthermore, Ap(7)A inhibits ADP-induced platelet aggregation. Thus, the potent vasoconstrictor Ap(7)A derived from human platelets, like other diadenosine polyphosphates, may play a role in the regulation of vascular tone and hemostasis.
AB - Diadenosine pentaphosphate and diadenosine hexaphosphate have been isolated in human platelets and have been postulated to play an important role in the control of vascular tone. Here we describe the isolation and identification of diadenosine heptaphosphate from human platelets. Dinucleoside polyphosphates were concentrated by affinity chromatography from a nucleotide-containing fraction from deproteinated human platelets. Dinucleoside polyphosphates were purified by anion-exchange and reversed phase high performance liquid chromatography to homogeneity. Analysis of one of these fractions with matrix-assisted laser desorption/ionization mass spectrometry revealed a molecular mass of 1076.4 (1077.4 = [M + H](+)) Da. UV spectroscopic analysis of this fraction showed the spectrum of an adenosine derivative. Comparison of the postsource decay matrix-assisted laser desorption/ionization mass spectrum of the fraction minus that of diadenosine heptaphosphate (Ap(7)A) demonstrated that the isolated substance was identical to Ap(7)A. The identity of the retention times of the authentic and the isolated compound confirmed this result. Enzymatic analysis demonstrated an interconnection of the phosphate groups with the adenosines in the 5'-positions of the riboses. With thrombin-induced platelet aggregation, Ap(7)A is released from the platelets into the extracellular space. The vasoconstrictive action of Ap(7)A on the vasculature of the isolated perfused rat kidney Ap(7)A was slightly less than that of Ap(6)A. The threshold of the vasoconstrictive action of Ap(7)A was 10(-5) mol/liter. The vasoconstrictive effect was abolished by suramin and pyridoxal phosphate 6-azophenyl-2', 4'-disulfonic acid, suggesting an activation of P(2x) receptors. Furthermore, Ap(7)A inhibits ADP-induced platelet aggregation. Thus, the potent vasoconstrictor Ap(7)A derived from human platelets, like other diadenosine polyphosphates, may play a role in the regulation of vascular tone and hemostasis.
KW - Adenosine Diphosphate
KW - Animals
KW - Blood Platelets
KW - Dinucleoside Phosphates
KW - Humans
KW - Platelet Aggregation
KW - Rats
KW - Suramin
KW - Thrombin
KW - Vasoconstriction
KW - Vasoconstrictor Agents
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1074/jbc.274.34.23926
DO - 10.1074/jbc.274.34.23926
M3 - SCORING: Journal article
C2 - 10446159
VL - 274
SP - 23926
EP - 23931
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 34
ER -