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ACS Chem. Biol. 1 (2),
83–91
10.1021/cb5000014
Web Release Date: March 3, 2006
Copyright © 2006 American Chemical Society
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Fluorogenic Phospholipids as Head Group-Selective Reporters of Phospholipase A Activity
Tyler M. Rose, and Glenn D. Prestwich*
Department of Medicinal Chemistry and Center for
Cell Signaling, University of Utah, 419 Wakara Way, Suite 205, Salt Lake
City, Utah 84108
Received for review November 11, 2005 and accepted January
27, 2006
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*To whom correspondence should be addressed. E-mail: Glenn.Prestwich#hsc.utah.edu.
PLA (phospholipases A) are important mediators of cell signaling, generating
bioactive fatty acids and LPLs (lysophospholipids). PLA products having different
head groups can initiate vastly different types of signaling. Fluorogenic analogues
of the PLs (phospholipids) PA (phosphatidic acid), PC (phosphatidylcholine),
PE (phosphatidylethanolamine), and PG (phosphatidylglycerol) were synthesized
as PLA substrates for rapidly determining in real time the influence of head
group modifications on cell signaling both in vitro and in cells. Enzyme-assisted
remodeling of the sn-2 position of the diacylglyceryl moiety with cobra
venom PLA2 and transphosphatidylation with a particular PLD (phospholipase
D) were central steps in the preparation of these enzymatic probes. The resulting
fluorogenic Dabcyl-and BODIPY-containing PL analogues, DBPA, DBPC, DBPE, and
DBPG, were used in mixed micelle assays to determine PLA2 kinetics.
Next, the assays were used to determine the Xi(50) value of
a common PLA2 inhibitor. Finally, the head group selectivities of
a series of commercially available PLA2 enzymes were readily established
using the DBPLs (Dabcyl-BODIPY PLs) as substrates.
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