Assignment of the heterogeneous static and time-resolved tryptophan fluorescence of 3-phosphoglycerate kinase.
The heterogeneous fluorescence of yeast 3-phosphoglycerate kinase, a hinge-bending enzyme with two tryptophan residues, has been resolved into three emission components using steady-state and time-resolved studies of the fluorescence quenching by acrylamide, iodide and caesium ions at different emission wavelengths. The buried Trp-333 has a blue-shifted heterogeneous emission spectrum characterised by three fluorescence lifetimes, and is inaccessible to quenchers. The surface Trp-308 also has a heterogeneous emission with multiple lifetimes. The emission of Trp-308 can be separated into a blue-shifted emission accessible to acrylamide and caesium only, and a red-shifted emission accessible to all three quenchers.
Dryden
D T
DT
Department of Biochemistry and Genetics, University of Newcastle upon Tyne, U.K.
Pain
R H
RH
D007700:Kinetics; D010735:Phosphoglycerate Kinase / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors; D014364:Tryptophan
D014364:Tryptophan; D010735:Phosphoglycerate Kinase