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Award Abstract #1264434

Analysis and Bioengineering of Optogenetic Rhodopsins

NSF Org: CBET
Div Of Chem, Bioeng, Env, & Transp Sys
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Initial Amendment Date: January 31, 2013
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Latest Amendment Date: January 31, 2013
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Award Number: 1264434
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Award Instrument: Standard Grant
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Program Manager: Leon Esterowitz
CBET Div Of Chem, Bioeng, Env, & Transp Sys
ENG Directorate For Engineering
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Start Date: July 1, 2013
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End Date: June 30, 2017 (Estimated)
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Awarded Amount to Date: $598,026.00
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Investigator(s): Kenneth Rothschild kjr@bu.edu (Principal Investigator)
Xue Han (Co-Principal Investigator)
Shyamsunder Erramilli (Co-Principal Investigator)
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Sponsor: Trustees of Boston University
881 COMMONWEALTH AVE
BOSTON, MA 02215-1300 (617)353-4365
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NSF Program(s): BIOPHOTONICS, IMAGING &SENSING
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Program Reference Code(s): 005E
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Program Element Code(s): 7236

ABSTRACT

1264434

Rothschild

The research will involve three laboratories at the BU Photonics Center (Rothschild, Erramilli and Han) and utilize advanced spectroscopic and biophysical/bioengineering techniques along with collaborators at UC Davis Center for Biophotonics, MIT, Harvard University, and University of Texas. A central feature of the research will be the use of time-resolved FTIR difference spectroscopic, ultra-fast laser spectroscopy and confocal near-IR Raman techniques to measure molecular changes in optogenetic rhodopsins upon light excitation on a time-scale of seconds to femtoseconds. Preliminary work has demonstrated that a variety of optogenetic rhodopsins can be reconstituted into proteolipid membranes including nanolipidparticles and studied using advanced spectroscopic methods to obtain detailed molecular information about their function. In addition, spectroscopic studies have been extended to living cells allowing these proteins to be studied in their native environment in order to determine molecule effects of such factors as membrane potential. Collaborators on this project include: Dr. Ed Boyden at MIT, widely recognized as one of the co-founders of optogenetics and a leading developer of new neurophotonic technology; Dr. Matt Coleman at UCD, who has developed methods of expressing rhodopsins into nanolipidparticles (NLPs) and forming microcrystals suitable for coherent x-ray diffraction imaging (CXDI), Dr. John Spudich, at the U. Texas Med. School, a pioneer in the field of microbial rhodopsins and co-discoverer of channelrhodopsins and Dr. Adam Cohen, whose laboratory at Harvard University has recently demonstrated that rhodopsins can be used to monitor transmembrane voltage in living nerves.


PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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Yi, A., Mamaeva, N., Li, H., Spudich, J. L., and Rothschild, K. J.. "Resonance Raman Study of an Anion Channelrhodopsin: Effects of Mutations near the Retinylidene Schiff Base.," Biochemistry, v.55, 2016, p. 2371. 

Ogren, J. I., Mamaev, S., Russano, D., Li, H., Spudich, J. L., and Rothschild, K. J.. "Retinal Chromophore Structure and Schiff Base Interactions in Red-Shifted Channelrhodopsin-1 from Chlamydomonas augustae," Biochemistry, 2014. 

Yi, A., Mamaeva, N., Li, H., Spudich, J. L., and Rothschild, K. J.. "Resonance Raman Study of an Anion Channelrhodopsin: Effects of Mutations near the Retinylidene Schiff Base," Biochemistry, v.55, 2016, p. 2371. 

Yi, A., Li, H., Mamaeva, N., Fernandez De Cordoba, R. E., Lugtenburg, J., DeGrip, W. J., Spudich, J. L., and Rothschild, K. J.. "Structural Changes in an Anion Channelrhodopsin: Formation of the K and L Intermediates at 80 K.," Biochemistry, v.56, 2017, p. 2197. 

Rothschild, K.J.. "The Early Development and Application of FTIR Difference Spectroscopy to Membrane Proteins: A Personal Perspective," Biomedical Spectroscopy and Imaging, 2016.

Ogren, J. I., Yi, A., Mamaev, S., Li, H., Spudich, J. L., and Rothschild, K. J.. "Proton Transfers in a Channelrhodopsin-1 Studied by FTIR-Difference Spectroscopy and Site-Directed Mutagenesis," Journal of Biological Chemistry, v.290, 2015, p. 12719. 

Casadei, Cecilia M. and Tsai, Ching-Ju and Barty, Anton and Hunter, Mark S. and Zatsepin, Nadia A. and Padeste, Celestino and Capitani, Guido and Benner, W. Henry and Boutet, Sébastien and Hau-Riege, Stefan P. and Kupitz, Christopher and Messerschmidt, Ma. "Resolution extension by image summing in serial femtosecond crystallography of two-dimensional membrane-protein crystals," IUCrJ, v.5, 2018.   

Ogren, J. I., Yi, A., Mamaev, S., Li, H., Lugtenburg, J., DeGrip, W. J., Spudich, J. L., and Rothschild, K. J.. "Comparison of the Structural Changes Occurring during the Primary Phototransition of Two Different Channelrhodopsins from Chlamydomonas Algae.," Biochemistry, v.54, 2015, p. 377. 

Ogren, J. I., Yi, A., Mamaev, S., Li, H., Spudich, J. L., and Rothschild, K. J.. "roton Transfers in a Channelrhodopsin-1 Studied by FTIR-Difference Spectroscopy and Site-Directed Mutagenesis," J Biol Chem, v.290, 2015, p. 12719. 

 

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