High-throughput mass spectrometry screening for inhibitors of phosphatidylserine decarboxylase.
A high-throughput mass spectrometry assay to measure the catalytic activity of phosphatidylserine decarboxylase (PISD) is described. PISD converts phosphatidylserine to phosphatidylethanolamine during lipid synthesis. Traditional methods of measuring PISD activity are low throughput and unsuitable for the high-throughput screening of large compound libraries. The high-throughput mass spectrometry assay directly measures phosphatidylserine and phosphatidylethanolamine using the RapidFiretrade mark platform at a rate of 1 sample every 7.5 s. The assay is robust, with an average Z' value of 0.79 from a screen of 9920 compounds. Of 60 compounds selected for confirmation, 54 are active in dose-response studies. The application of high-throughput mass spectrometry permitted a high-quality screen to be performed for an otherwise intractable target.
Forbes
Chris D
CD
Department of Research Technologies, Bayer Pharmaceuticals Corporation, West Haven, CT 06516, USA.
Toth
Joshuaine G
JG
Ozbal
Can C
CC
Lamarr
William A
WA
Pendleton
Jennifer A
JA
Rocks
Sandra
S
Gedrich
Richard W
RW
Osterman
David G
DG
Landro
James A
JA
Lumb
Kevin J
KJ
D002262:Carboxy-Lyases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000235:genetics; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004795:Enzyme Stability; D005615:Freezing; D006801:Humans; D007668:Kidney / Q000166:cytology; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors; D012371:Robotics; D017422:Sequence Analysis, DNA; D014162:Transfection
D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase