Single-molecule fluorescence spectroscopy of enzyme conformational dynamics and cleavage mechanism

Taekjip Ha, Alice Y. Ting, Joy Liang, W. Brett Caldwell, Ashok A. Deniz, Daniel S. Chemla, Peter G. Schultz, Shimon Weiss

Research output: Contribution to journalArticlepeer-review

Abstract

Fluorescence resonance energy transfer and fluorescence polarization anisotropy are used to investigate single molecules of the enzyme staphylococcal nuclease. Intramolecular fluorescence resonance energy transfer and fluorescence polarization anisotropy measurements of fluorescently labeled staphylococcal nuclease molecules reveal distinct patterns of fluctuations that may be attributed to protein conformational dynamics on the millisecond time scale. Intermolecular fluorescence resonance energy transfer measurements provide information about the dynamic interactions of staphylococcal nuclease with single substrate molecules. The experimental methods demonstrated here should prove generally useful in studies of protein folding and enzyme catalysis at single-molecule resolution.

Original languageEnglish (US)
Pages (from-to)893-898
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number3
DOIs
StatePublished - Feb 2 1999
Externally publishedYes

ASJC Scopus subject areas

  • General

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