High-throughput screen identifies cyclic nucleotide analogs that inhibit prostatic acid phosphatase.
High-throughput screen identifies cyclic nucleotide analogs that inhibit prostatic acid phosphatase.
复制标题
DOI：
10.1177/1087057112468613
复制
发表时间：
2013-04
影响因子：
--
通讯作者：
Zylka MJ
中科院分区：
文献类型：
--
作者：
McCoy ES;Lea WA;Mott BT;Maloney DJ;Jadhav A;Simeonov A;Zylka MJ
The secretory and transmembrane isoforms of Prostatic acid phosphatase (PAP) can dephosphorylate extracellular adenosine 5′-monophosphate (AMP) to adenosine, classifying PAP as an ectonucleotidase. Currently, there are no compounds that inhibit PAP in living cells. To identify small molecule modulators of PAP, we used a 1,536-well based quantitative high-throughput fluorogenic assay to screen the Library of Pharmacologically Active Compounds (LOPAC1280) arrayed as eight-concentration dilution series. This fluorogenic assay used difluoro-4-methylumbelliferyl phosphate (DiFMUP) as substrate and collected data in kinetic mode. Candidate hits were subsequently tested in an orthogonal absorbance-based biochemical assay that used AMP as substrate. From these initial screens, three inhibitors of secretory human (h) and mouse (m)PAP were identified: 8-(4-chlorophenylthio) cAMP (pCPT-cAMP), calmidazolium chloride and nalidixic acid. These compounds did not inhibit recombinant alkaline phosphatase. Of these compounds, only pCPT-cAMP and a related cyclic nucleotide analog [8-(4-chlorophenylthio) cGMP; pCPT-cGMP] inhibited the ectonucleotidase activity of transmembrane PAP in a cell-based assay. These cyclic nucleotides are structurally similar to AMP but cannot be hydrolyzed by PAP. In summary, we identified two cyclic nucleotide analogs that inhibit secretory and transmembrane PAP in vitro and in live cells.
登录
查看更多内容
DOI：
10.1016/0020-711x(91)90047-q
发表时间：
1991-01-01
期刊：
INTERNATIONAL JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
DAI, XY;SNOW, LD
通讯作者：
SNOW, LD
影响因子：
5
作者：
OSTROWSKI, WS;KUCIEL, R
通讯作者：
KUCIEL, R
影响因子：
--
作者：
Zhang, JH;Chung, TDY;Oldenburg, KR
通讯作者：
Oldenburg, KR
影响因子：
11.2
作者：
Quintero, Ileana B.;Araujo, Cesar L.;Vihko, Pirkko T.
通讯作者：
Vihko, Pirkko T.
影响因子：
4.8
作者：
Waidmann, Oliver;Pleli, Thomas;Piiper, Albrecht
通讯作者：
Piiper, Albrecht