Time-resolved structure determinations are at the forefront of attempts to understand the mechanisms of protein reactions. We have demonstrated a system for collecting high quality x-ray absorption spectroscopy data on microsecond timescales. The method can be used for both full spectra accumulation and kinetic spectro-photometry. These experiments have vast potential, not only for examining metalloproteins, but also for observing the dynamic behavior of a wide range of chemical elements in conjunction with photochemical, rapid mixing, and temperature jump techniques. The following accomplishments have been made in the preceeding 12-month period: 1) We have successfully increased the effective X-ray flux more than 10 fold by implementing the sagittal dynamic focusing monochromator. 2) We have increased the signal-to-noise ratio by nearly 3 times, with the design, construction and reliable operation of time-resolved 14 channel window discriminator copuled to a computer controlled data acquisition system. 3) We have permanently installed a 20 Hz Nd-YAG laser system with 532 and 355 nm pump beams on beamline X-9B for initiating time-resolved photo-reactions. 4) We have published two papers detailing these improvements, along with several published abstracts and meeting presentations and have another manuscript in preparation.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001633-14
Application #
5223455
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
14
Fiscal Year
1996
Total Cost
Indirect Cost
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Masip, Lluis; Pan, Jonathan L; Haldar, Suranjana et al. (2004) An engineered pathway for the formation of protein disulfide bonds. Science 303:1185-9
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Swisher, Jennifer F; Su, Linhui J; Brenowitz, Michael et al. (2002) Productive folding to the native state by a group II intron ribozyme. J Mol Biol 315:297-310
Dhavan, Gauri M; Crothers, Donald M; Chance, Mark R et al. (2002) Concerted binding and bending of DNA by Escherichia coli integration host factor. J Mol Biol 315:1027-37

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