Abstract

The dynamical behavior of proteins will be investigated, using time-resolved Raman and infrared spectroscopies as probes of structure in real time. Laser temperature-jump techniques will be applied to the protein folding problem in order to characterize the earliest stages in folding and unfolding. T-jump studies will be extended to investigate protein motions tied to function: enzyme activity in alcohol dehydrogenase, iron release from ferritin, and apoptosis triggering by cytochrome c. The dynamics of protein allostery will be investigated, using hemoglobin as a paradigm. Pump-probe spectroscopic monitoring of the R-T conformation change will be used to define the reaction coordinate in molecular detail, using site-directed mutagenesis and metal substitution, in conjunction with computational modeling. By increasing our understanding of hemoglobin, these studies may lead to improved hematological therapies.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM025158-33
Application #
7879225
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Smith, Ward
Project Start
1978-06-01
Project End
2011-09-14
Budget Start
2010-07-01
Budget End
2011-09-14
Support Year
33
Fiscal Year
2010
Total Cost
$330,502
Indirect Cost

  Institution

Name
University of Washington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Vaughn, Morgan B; Zhang, Jianyu; Spiro, Thomas G et al. (2018) Activity-Related Microsecond Dynamics Revealed by Temperature-Jump Förster Resonance Energy Transfer Measurements on Thermophilic Alcohol Dehydrogenase. J Am Chem Soc 140:900-903
Meadows, Corey W; Balakrishnan, Gurusamy; Kier, Brandon L et al. (2015) Temperature-Jump Fluorescence Provides Evidence for Fully Reversible Microsecond Dynamics in a Thermophilic Alcohol Dehydrogenase. J Am Chem Soc 137:10060-3
Jones, Eric M; Balakrishnan, Gurusamy; Squier, Thomas C et al. (2014) Distinguishing unfolding and functional conformational transitions of calmodulin using ultraviolet resonance Raman spectroscopy. Protein Sci 23:1094-101
Balakrishnan, Gurusamy; Soldatova, Alexandra V; Reid, Philip J et al. (2014) Ultrafast charge transfer in nickel phthalocyanine probed by femtosecond Raman-induced Kerr effect spectroscopy. J Am Chem Soc 136:8746-54
Jones, Eric M; Monza, Emanuele; Balakrishnan, Gurusamy et al. (2014) Differential control of heme reactivity in alpha and beta subunits of hemoglobin: a combined Raman spectroscopic and computational study. J Am Chem Soc 136:10325-39
Soldatova, Alexandra V; Ibrahim, Mohammed; Spiro, Thomas G (2013) Electronic structure and ligand vibrations in FeNO, CoNO, and FeOO porphyrin adducts. Inorg Chem 52:7478-86
Spiro, Thomas G; Soldatova, Alexandra V; Balakrishnan, Gurusamy (2013) CO, NO and O2 as Vibrational Probes of Heme Protein Interactions. Coord Chem Rev 257:511-527
Balakrishnan, Gurusamy; Hu, Ying; Spiro, Thomas G (2012) His26 protonation in cytochrome c triggers microsecond ?-sheet formation and heme exposure: implications for apoptosis. J Am Chem Soc 134:19061-9
Balakrishnan, Gurusamy; Jarzecki, Andrzej A; Wu, Qiang et al. (2012) Mode recognition in UV resonance Raman spectra of imidazole: histidine monitoring in proteins. J Phys Chem B 116:9387-95
Jones, Eric M; Balakrishnan, Gurusamy; Spiro, Thomas G (2012) Heme reactivity is uncoupled from quaternary structure in gel-encapsulated hemoglobin: a resonance Raman spectroscopic study. J Am Chem Soc 134:3461-71

Showing the most recent 10 out of 61 publications