Abstract
Fluorogenic analogues of phosphatidylcholine and lysophosphatidylcholine, DDPB and lysoDDPB, were synthesized by an enzyme-assisted strategy. The analogues were evaluated as substrates for phospholipases C and D and lysophospholipase D. DDPB was cleaved by bacterial and plant phospholipase D (PLD) enzymes and represents the first direct fluorogenic substrate for real-time measurement of PLD activity. Both fluorogenic substrates have potential in screening for PLD and PC-PLC inhibitors and for monitoring spatiotemporal changes in PLD activity in cells. [structure: see text]
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Bacillus cereus / enzymology
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Clostridium perfringens / enzymology
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Enzyme Activation / physiology
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Fluorescent Dyes / chemical synthesis*
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Fluorescent Dyes / metabolism
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Lysophosphatidylcholines / chemical synthesis*
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Lysophosphatidylcholines / metabolism
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Phosphatidylcholines / chemical synthesis*
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Phosphatidylcholines / metabolism
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Phospholipase D / chemistry*
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Phospholipase D / metabolism*
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Phosphoric Diester Hydrolases / metabolism
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Spider Venoms / enzymology
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Spider Venoms / metabolism
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Type C Phospholipases / chemistry*
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Type C Phospholipases / metabolism*
Substances
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Fluorescent Dyes
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Lysophosphatidylcholines
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Phosphatidylcholines
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Spider Venoms
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loxosceles venom
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Phosphoric Diester Hydrolases
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Type C Phospholipases
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Phospholipase D