Monday, 23 May 2005

This presentation is part of: Spectroscopy of Biomolecules Posters

Single Molecule Measurement of Fast Folding Proteins Using Fluorescence Resonance Energy Transfer Confocal Microscopy

Jongjin Jung, Hiyun Kim, Troy C. Messina, Jason T. Giurleo, and David S. Talaga. Rutgers University, Piscataway, NJ

We are studying the folding dynamics of a l85 repressor, the truncated N-terminal domain of l repressor, using single molecule measurements. The l85 repressor with double mutants (Gly46 Ala, Gly48 Ala) was reported to fold with a lifetime of less than 20ms (RE. Burton et al J.Mol.Biol 1996 263 311- 322) without intermediate states. The mutant protein, G46A/G48A, has been labeled with a fluorescent donor (Alexa488) and acceptor (Cy7) at its N and C termini. Using a Fluorescence Resonance Energy Transfer (FRET) measurement, we can investigate the dynamics of fast-folding proteins in two different environments, i) one in which a protein is confined but freely rotating in agarose gel, ii) the other in which a protein is fixated on glass surface through nitrilotriacetic acid (NTA) - chelated Nikel. In addition to the end-to-end distance distributions by the different urea concentrations, the instantaneous events during folding in a microsecond time scale and the fast-folding dynamics will be elucidated to explore the folding landscape and the memory effects of the G46A/G48A by applying novel single molecule spectroscopy analysis tool, Hidden Markov Model (HMM).

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