Understanding how various biomolecules embedded within biochemical and mechanical networks are coordinated over multiple length and time scales to produce orchestrated cell behaviors requires characterization of their biochemical and biophysical properties in their native cellular context. However this represents a major challenge and our current understanding of the native behaviors of biomolecules has been limited by the lack of suitable techniques to measure and manipulate their dynamic properties in living cells. For example, to reveal causal connections and achieve a systems understanding of the signal transduction networks in living cells, a large set of molecular tools, designed to specifically perturb signaling pathways in situ and to quantitatively measure the cellular response, must be developed. Furthermore, very little is known about the mechanical properties and behaviors of molecules involved in force generation, cellular mechanics maintenance and mechanotransduction in a living cell due, in large part, to the lack of experimental methodologies for intracellular measurement and manipulation of biophysical properties. I propose to develop such molecular tools as genetically encodable magnetic probes by integrating molecular engineering strategies, chemical and cell biological approaches, and state-of-the-art biophysical measurement and manipulation combining single-molecule imaging and magnetic tweezers. This work will identify new strategies for simultaneous measurement and manipulation of intracellular molecular events and will provide enabling technologies for probing biochemical and biophysical processes in living cells. A better understanding of life processes at the molecular level will lead to new insights in treating diseases that result from dysregulation of these processes.

Public Health Relevance

The goal of this application is to develop novel magnetic probes that can be genetically encoded to enable new ways of measuring and manipulating the biochemical and biophysical properties of macromolecules in living cells. Once developed, these probes should serve as powerful tools for understanding fundamental life processes, which in turn facilitates treatment of diseases that result from dysregulation of these processes.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
NIH Director’s Pioneer Award (NDPA) (DP1)
Project #
5DP1CA174423-05
Application #
8522266
Study Section
Special Emphasis Panel ()
Program Officer
Knowlton, John R
Project Start
2009-09-30
Project End
2015-07-31
Budget Start
2014-08-01
Budget End
2015-07-31
Support Year
5
Fiscal Year
2014
Total Cost
$799,623
Indirect Cost
$312,048
Name
Johns Hopkins University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
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Zhou, Xin; Li, Simin; Zhang, Jin (2016) Tracking the Activity of mTORC1 in Living Cells Using Genetically Encoded FRET-based Biosensor TORCAR. Curr Protoc Chem Biol 8:225-233
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Mehta, Sohum; Zhang, Jin (2015) Dynamic visualization of calcium-dependent signaling in cellular microdomains. Cell Calcium 58:333-41
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Zhou, Xin; Clister, Terri L; Lowry, Pamela R et al. (2015) Dynamic Visualization of mTORC1 Activity in Living Cells. Cell Rep :
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Hu, Jianfei; Neiswinger, Johnathan; Zhang, Jin et al. (2015) Systematic Prediction of Scaffold Proteins Reveals New Design Principles in Scaffold-Mediated Signal Transduction. PLoS Comput Biol 11:e1004508

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