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Projects 2005

Sponsoring Laboratory/Section:

Laboratory of Biophysical Chemistry, Optical Spectroscopy Section
National Heart, Lung and Blood Institute (NHLBI)

Lab Chief:

James Ferretti, Ph.D.

Mentors:

Jay R. Knutson, Alex Smirnov, Jianhua Xu,
Kristi Wojtuszewski Poulin

Contact Information:

e-mail: jaysan@helix.nih.gov
phone: (301) 496-2557
fax: (301) 480-6964

Laboratory Description and Project area:

We develop and exploit the techniques of laser-driven, time-resolved fluorescence spectroscopy, especially in protein-DNA and two-photon microscopy. We develop state-of-the art ultrafast laser instruments that help us determine the local flexibility, size and structure of complexes in solution. For example, we have been studying HIV integrase, the enzyme that incorporates the AIDS virus into human DNA, and we are learning about the stoichiometry and shape of the active complex. Recently, we have been developing both 2-photon lifetime and FCS microscopy methods to extend the spectroscopic approach to the cellular level. We have, for example, examined the free/bound ratio of NADH in cardiac mitochondria. We also develop the technique itself- building new upconversion fluorometers based on femtosecond lasers, new stopped-flow lifetime instruments and new optimization (eg., genetic) algorithms for data analysis. The summer interns will, depending on their interests,

• be introduced to fluorescence instruments and data acquisition;
• help develop computer software to analyze global structure features and flexibility of proteins and DNA;
• help develop robotic control of novel microscopes or fluorometers;
• help develop femtosecond laser instrumentation; or
• learn site-specific mutant protein preparation.