Fluorescence quenched quinone methide based activity probes--a cautionary tale.
A carbamate linked quenching group coupled with a pro-quinone methide reactive core provides an effective tool for studying enzyme function without problems associated with background fluorescence from unreacted probe. However, the relatively slow fragmentation of the carbamate linkage in such a strategy may cause problems of loss of signal or a decoupling of enzyme activity and labelling.
Centre for Bioactive Chemistry, Department of Chemistry, University of Durham, Sciences Laboratories, South Road, Durham, UK DH1 3LE.
Jonathan D
JD
Sellars
Marie
M
Landrum
Aileen
A
Congreve
David P
DP
Dixon
Jackie A
JA
Mosely
Andrew
A
Beeby
Robert
R
Edwards
Patrick G
PG
Steel
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D002219:Carbamates / Q000737:chemistry*; D043182:Carboxylesterase / Q000032:analysis*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D045563:Indolequinones / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016415:Sequence Alignment
D002219:Carbamates; D005456:Fluorescent Dyes; D045563:Indolequinones; C068040:quinone methide; D043182:Carboxylesterase