Lu Xuequan, Cseh Sandor, Byun Hoe-Sup, Tigyi Gabor, Bittman Robert
Department of Chemistry and Biochemistry, Queens College of The City University of New York, Flushing, New York 11367-1597, USA.
J Org Chem. 2003 Sep 5;68(18):7046-50. doi: 10.1021/jo034828q.
The first synthesis of two photoreactive analogues of the lipid mediator and second messenger sphingosine 1-phosphate (S1P), [(32)P]-labeled (2S,3R)-14-O-(4'-benzoylphenyl)- and (2S,3R)-14-O-((4'-trifluoromethyldiazirinyl)phenyl)-(4E)-tetradecenyl-2-amino-3-hydroxy-1-phosphate, is described. The interactions of these probes with the S1P type-1 receptor (S1P(1)) transfected into membranes of rat hepatoma cells and with plasma proteins were analyzed. The S1P(1) receptor interacted in a specific manner with the benzophenone-containing ligand (K(D) = 84 +/- 10 nM vs K(D) for S1P = 36 +/- 2 nM); in contrast, no saturable specific binding was found with the diazirine-containing ligand. However, the same pattern was found for labeling of plasma proteins by both probes, indicating that different parts of the S1P pharmacophore underlie the interaction of S1P with its receptor and plasma carrier proteins.
本文描述了脂质介质和第二信使鞘氨醇-1-磷酸（S1P）的两种光反应性类似物的首次合成，即[(32)P]标记的(2S,3R)-14-O-(4'-苯甲酰基苯基)-和(2S,3R)-14-O-((4'-三氟甲基二氮杂环丁烷基)phenyl)-(4E)-十四碳烯基-2-氨基-3-羟基-1-磷酸。分析了这些探针与转染到大鼠肝癌细胞膜中的S1P 1型受体（S1P(1)）以及与血浆蛋白的相互作用。S1P(1)受体以特定方式与含二苯甲酮的配体相互作用（K(D) = 84 +/- 10 nM，而S1P的K(D) = 36 +/- 2 nM）；相比之下，含二氮杂环丁烷的配体未发现可饱和的特异性结合。然而，两种探针标记血浆蛋白的模式相同，表明S1P药效基团的不同部分是S1P与其受体和血浆载体蛋白相互作用的基础。
