Bovine serum albumin-(7-hydroxycoumarin-4-acetic acid) complex: applications to the fluorometric measurement of fatty acid concentrations.
A covalent complex between bovine serum albumin and 7-hydroxycoumarin-4-acetic acid (BSA-HCA) shows a strong fluorescence band at lambdamax = 450 nm upon excitation at 375 nm. Quenching of the fluorescence emission accompanies the association of fatty acids (FA) to BSA-HCA and the application of the complex as a spectrofluorometric probe for measurement of fatty acid concentrations in aqueous solution is examined. Binding constants for various long-chain fatty acids (Kd = 14-460 nM) and calibration curves characterizing the probe have been determined. Standardized assay conditions allow for accurate measurements in the concentration range of 10 nM to 5 microM. BSA-HCA provides a stable and sensitive fluorescence-based FA probe with potential biochemical applications.
Demant
E J
EJ
Biochemistry Laboratory C, Panum Institute, University of Copenhagen, Copenhagen, DK-2200, Denmark.
D000085:Acetates / Q000378:metabolism*; D003374:Coumarins / Q000378:metabolism*; D005227:Fatty Acids / Q000032:analysis* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism*; D005470:Fluorometry; D012710:Serum Albumin, Bovine / Q000378:metabolism*
D000085:Acetates; D003374:Coumarins; D005227:Fatty Acids; D005456:Fluorescent Dyes; D012710:Serum Albumin, Bovine; C102522:7-hydroxycoumarin-4-acetic acid