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 #
5R01GM025158-31
Application #
7440264
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Smith, Ward
Project Start
1978-06-01
Project End
2011-06-30
Budget Start
2008-07-01
Budget End
2009-06-30
Support Year
31
Fiscal Year
2008
Total Cost
$333,840
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