TY - JOUR
T1 - Submicrosecond real-time fluorescence sampling
T2 - Application to protein folding
AU - Ervin, John
AU - Sabelko, Jobiah
AU - Gruebele, Martin
N1 - Funding Information:
This project was funded by NIH grant GM057175 and a Lucile and David Packard Fellowship. We thank Professor Doug McDonald for helpful discussions about the dispersed fluorescence detection. The ubiquitin expression system was a gift of Professor Tracy Handel at the University of California at Berkeley; the PGK expression system was obtained from Professor Maria Mas and Genentech. M.G. was an Alfred P. Sloan Foundation Fellow and Dreyfus Teacher-Scholar while this project was carried out.
PY - 2000/1/30
Y1 - 2000/1/30
N2 - Time-resolved fluorescence detection has become a central tool in the study of protein folding. This article briefly reviews modern fluorescence techniques and then focuses on recent improvements made possible by array photomultipliers, computer-controlled data gating, and long-memory multi-channel digitizers. It is now possible to detect fluorescence wavelength profiles and/or fluorescence decay transients very cost effectively with sub-microsecond kinetic time resolution out to long times. Folding kinetics can be analyzed by singular value decomposition (SVD) or χ-analysis. The latter provides an objective method for detecting nonexponential kinetics in two-state systems. (C) 2000 Elsevier Science S.A.
AB - Time-resolved fluorescence detection has become a central tool in the study of protein folding. This article briefly reviews modern fluorescence techniques and then focuses on recent improvements made possible by array photomultipliers, computer-controlled data gating, and long-memory multi-channel digitizers. It is now possible to detect fluorescence wavelength profiles and/or fluorescence decay transients very cost effectively with sub-microsecond kinetic time resolution out to long times. Folding kinetics can be analyzed by singular value decomposition (SVD) or χ-analysis. The latter provides an objective method for detecting nonexponential kinetics in two-state systems. (C) 2000 Elsevier Science S.A.
KW - Apomyoglobin
KW - Nonexponential kinetics
KW - Phosphoglycerate kinase
KW - Tryptophan
KW - Ubiquitin
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U2 - 10.1016/S1011-1344(00)00002-6
DO - 10.1016/S1011-1344(00)00002-6
M3 - Review article
C2 - 10739138
AN - SCOPUS:0033996482
SN - 1011-1344
VL - 54
SP - 1
EP - 15
JO - Journal of Photochemistry and Photobiology B: Biology
JF - Journal of Photochemistry and Photobiology B: Biology
IS - 1
ER -