Aldol sensors for the rapid generation of tunable fluorescence by antibody catalysis.
The synthesis of novel fluorogenic retro-aldol substrates for aldolase antibody 38C2 is described. These substrates are efficiently and specifically processed by antibody aldolases but not by natural cellular enzymes. Together, the fluorogenic substrates and antibody aldolases provide reporter gene systems that are compatible with living cells. The broad scope of the antibody aldolase allows for the processing of a range of substrates that can be designed to allow fluorescence monitoring at a variety of wavelengths. We also have developed the following concept in fluorescent protein tags. beta-Diketones bearing a fluorescent tag are bound covalently by the aldolase antibody and not other proteins. We anticipate that proteins fused with the antibody can be tagged specifically and covalently within living cells with fluorophores of virtually any color, thereby providing an alternative to green fluorescent protein fusions.
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The Skaggs Institute for Chemical Biology and the Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Barbas
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Lerner
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D000447:Aldehydes / Q000138:chemical synthesis* / Q000737:chemistry; D017151:Antibodies, Catalytic / Q000378:metabolism*; D015374:Biosensing Techniques; D002384:Catalysis; D005456:Fluorescent Dyes; D005634:Fructose-Bisphosphate Aldolase / Q000378:metabolism*; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism*; D007202:Indicators and Reagents; D007700:Kinetics; D015394:Molecular Structure; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D000447:Aldehydes; D017151:Antibodies, Catalytic; D005456:Fluorescent Dyes; D007140:Immunoglobulin Fab Fragments; D007202:Indicators and Reagents; C116609:3-hydroxybutanal; C109536:antibody aldolase; D005634:Fructose-Bisphosphate Aldolase