Submicrosecond real-time fluorescence sampling: Application to protein folding

John Ervin; Jobiah Sabelko; Martin Gruebele

doi:10.1016/S1011-1344(00)00002-6

Submicrosecond real-time fluorescence sampling: Application to protein folding

John Ervin, Jobiah Sabelko, Martin Gruebele

Research output: Contribution to journal › Review article › peer-review

Abstract

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.

Original language	English (US)
Pages (from-to)	1-15
Number of pages	15
Journal	Journal of Photochemistry and Photobiology B: Biology
Volume	54
Issue number	1
DOIs	https://doi.org/10.1016/S1011-1344(00)00002-6
State	Published - Jan 30 2000

Keywords

Apomyoglobin
Nonexponential kinetics
Phosphoglycerate kinase
Tryptophan
Ubiquitin

ASJC Scopus subject areas

Radiation
Radiological and Ultrasound Technology
Biophysics
Radiology Nuclear Medicine and imaging

Online availability

10.1016/S1011-1344(00)00002-6

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title = "Submicrosecond real-time fluorescence sampling: Application to protein folding",

abstract = "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.",

keywords = "Apomyoglobin, Nonexponential kinetics, Phosphoglycerate kinase, Tryptophan, Ubiquitin",

author = "John Ervin and Jobiah Sabelko and Martin Gruebele",

note = "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.",

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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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Submicrosecond real-time fluorescence sampling: Application to protein folding

Abstract

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