TY - JOUR
T1 - Subtype specific internalization of P2Y 1 and P2Y 2 receptors induced by novel adenosine 5′-O-(1-boranotriphosphate) derivatives
AU - Tulapurkar, M. E.
AU - Laubinger, W.
AU - Nahum, V.
AU - Fischer, B.
AU - Reiser, G.
PY - 2004/7
Y1 - 2004/7
N2 - 1. P2Y-nucleotide receptors represent important targets for drug development. The lack of stable and receptor specific agonists, however, has prevented successful therapeutic applications. A novel series of P-boronated ATP derivatives (ATP-α-B) were synthesized by substitution of a nonbridging O at P α with a BH 3 group. This introduces a chiral center, thus resulting in diastereoisomers. In addition, at C2 of the adenine ring a further substitution was made (Cl- or methylthio-). The pairs of diastereoisomers were denoted here as A and B isomers. 2. Here, we tested the receptor subtype specificity of these analogs on HEK 293 cells stably expressing rat P2Y 1 and rat P2Y 2 receptors, respectively, both attached to the fluorescent marker protein GFP (rP2Y 1-GFP, rP2Y 2-GFP). We investigated agonist-induced receptor endocytosis, [Ca 2+] i rise and arachidonic acid (AA) release. 3. Agonist-induced endocytosis of rP2Y 1-GFP was more pronounced for the A isomers than the corresponding B counterparts for all ATP-α-B analogs. Both 2-MeS-substituted diastereoisomers induced a greater degree of agonist-induced receptor endocytosis as compared to the 2-Cl-substituted derivatives. Endocytosis results are in accordance with the potency to induce Ca 2+ release by these compounds in HEK 293 cells stably transfected with rP2Y 1. 4. In case of rP2Y 2-GFP, the borano-nucleotides were very weak agonists in comparison to UTP and ATP in terms of Ca 2+ release, AA release and in inducing receptor endocytosis. The different ATP-α-B derivatives and also the diastereoisomers were equally ineffective. 5. Thus, the new agonists may be considered as potent and highly specific agonist drug candidates for P2Y 1 receptors. The difference in activity of the ATP analogs at P2Y receptors could be used as a tool to investigate structural differences between P2Y receptor subtypes.
AB - 1. P2Y-nucleotide receptors represent important targets for drug development. The lack of stable and receptor specific agonists, however, has prevented successful therapeutic applications. A novel series of P-boronated ATP derivatives (ATP-α-B) were synthesized by substitution of a nonbridging O at P α with a BH 3 group. This introduces a chiral center, thus resulting in diastereoisomers. In addition, at C2 of the adenine ring a further substitution was made (Cl- or methylthio-). The pairs of diastereoisomers were denoted here as A and B isomers. 2. Here, we tested the receptor subtype specificity of these analogs on HEK 293 cells stably expressing rat P2Y 1 and rat P2Y 2 receptors, respectively, both attached to the fluorescent marker protein GFP (rP2Y 1-GFP, rP2Y 2-GFP). We investigated agonist-induced receptor endocytosis, [Ca 2+] i rise and arachidonic acid (AA) release. 3. Agonist-induced endocytosis of rP2Y 1-GFP was more pronounced for the A isomers than the corresponding B counterparts for all ATP-α-B analogs. Both 2-MeS-substituted diastereoisomers induced a greater degree of agonist-induced receptor endocytosis as compared to the 2-Cl-substituted derivatives. Endocytosis results are in accordance with the potency to induce Ca 2+ release by these compounds in HEK 293 cells stably transfected with rP2Y 1. 4. In case of rP2Y 2-GFP, the borano-nucleotides were very weak agonists in comparison to UTP and ATP in terms of Ca 2+ release, AA release and in inducing receptor endocytosis. The different ATP-α-B derivatives and also the diastereoisomers were equally ineffective. 5. Thus, the new agonists may be considered as potent and highly specific agonist drug candidates for P2Y 1 receptors. The difference in activity of the ATP analogs at P2Y receptors could be used as a tool to investigate structural differences between P2Y receptor subtypes.
KW - Adenosine 5′-O-(1-boranotriphosphate) derivatives
KW - Green fluorescent protein
KW - Human embryonic kidney (HEK 293) cells
KW - Purinergic receptor
KW - Receptor endocytosis
UR - http://www.scopus.com/inward/record.url?scp=3242884617&partnerID=8YFLogxK
U2 - 10.1038/sj.bjp.0705859
DO - 10.1038/sj.bjp.0705859
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C2 - 15197109
AN - SCOPUS:3242884617
SN - 0007-1188
VL - 142
SP - 869
EP - 878
JO - British Journal of Pharmacology
JF - British Journal of Pharmacology
IS - 5
ER -