A green fluorescent chemosensor for amino acids provides a versatile high-throughput screening (HTS) assay for proteases.
The water soluble fluorescein-based ligand 1 forms a non-fluorescent complex with Cu(2+). This complex serves as a fluorescent sensor for amino acids in the 10(-3) M concentration range. Since the signal response is very fast, the sensor can be used to detect the hydrolytic activity of various proteases (trypsin, chymotrypsin, subtilisin) on bovine serum albumin as a whole protein substrate, and more generally to follow reactions releasing or removing free amino acids, in real time.
Dean
Kathryn E S
KE
Department of Chemistry & Biochemistry, University of Bern, Freiestrasse 3, Switzerland.
Klein
Gérard
G
Renaudet
Olivier
O
Reymond
Jean-Louis
JL
D000596:Amino Acids / Q000032:analysis*; D003300:Copper; D004353:Drug Evaluation, Preclinical; D010450:Endopeptidases / Q000378:metabolism*; D005452:Fluoresceins; D005453:Fluorescence; D007700:Kinetics; D008024:Ligands; D015335:Molecular Probes / Q000737:chemistry*
D000596:Amino Acids; D005452:Fluoresceins; D008024:Ligands; D015335:Molecular Probes; D003300:Copper; D010450:Endopeptidases; C007740:fluorexon