The synthesis of new proteins is fundamentally linked with learning and memory. At the molecular level there is strong evidence linking specific types of synaptic activity with the local synthesis of new protein. This grant application proposes the creation of new photochemical tools that will enable researchers to use light to manipulate translation and thereby control where and when proteins are made in neuronal cell culture, in brain slices and ultimately in living animals. The approach taken to the creation of these tools will be chemical synthesis based on the known structures of key components of the biochemical pathways that produce new proteins. Light- activated versions of well-known protein synthesis inhibitors (puromycin, rapamycin, 4E binding proteins) will be created, including reversible versions based on azobenzene photo-switches that can be turned on with light and that turn off in the dark. A second generation of light-activated inhibitors based on photo-active yellow protein/4E binding protein fusions is planned that is genetically-encoded. Genetically-encoded inhibitors can be selectively expressed in particular cells thereby enabling well-defined manipulation of the molecular events involved in learning and memory in whole living animals. Use of these tools thus promises to help uncover how learning and memory occur in living animals at a molecular level.

Public Health Relevance

Numerous mental health problems including post-traumatic stress disorder, epilepsy, obsessive compulsive disorders, or addiction are connected with "mis-wiring" of the brain, i.e. aberrant synaptic plasticity. Understanding how this works at the molecular level will show the path to effective treatment.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH086379-04
Application #
8298472
Study Section
Special Emphasis Panel (ZRG1-ETTN-G (52))
Program Officer
Freund, Michelle
Project Start
2009-09-30
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2014-05-31
Support Year
4
Fiscal Year
2012
Total Cost
$163,814
Indirect Cost
$12,134
Name
University of Toronto
Department
Type
DUNS #
259999779
City
Toronto
State
ON
Country
Canada
Zip Code
M5 1-S8
Reis, Jakeb M; Burns, Darcy C; Woolley, G Andrew (2014) Optical control of protein-protein interactions via blue light-induced domain swapping. Biochemistry 53:5008-16
Samanta, Subhas; Beharry, Andrew A; Sadovski, Oleg et al. (2013) Photoswitching azo compounds in vivo with red light. J Am Chem Soc 135:9777-84
Samanta, Subhas; Babalhavaeji, Amirhossein; Dong, Ming-xin et al. (2013) Photoswitching of ortho-substituted azonium ions by red light in whole blood. Angew Chem Int Ed Engl 52:14127-30
Kumar, Anil; Burns, Darcy C; Al-Abdul-Wahid, M Sameer et al. (2013) A circularly permuted photoactive yellow protein as a scaffold for photoswitch design. Biochemistry 52:3320-31
Beharry, Andrew A; Woolley, G Andrew (2011) Azobenzene photoswitches for biomolecules. Chem Soc Rev 40:4422-37
Fan, Helen Y; Morgan, Stacy-Anne; Brechun, Katherine E et al. (2011) Improving a designed photocontrolled DNA-binding protein. Biochemistry 50:1226-37
Samanta, Subhas; Woolley, G Andrew (2011) Bis-azobenzene crosslinkers for photocontrol of peptide structure. Chembiochem 12:1712-23
Sadovski, Oleg; Jaikaran, Anna S I; Samanta, Subhas et al. (2010) A collection of caged compounds for probing roles of local translation in neurobiology. Bioorg Med Chem 18:7746-52