Document Detail

Orientation-based FRET sensor for real-time imaging of cellular forces.
MedLine Citation:
PMID:  22389408     Owner:  NLM     Status:  MEDLINE    
Mechanical stress is an unmapped source of free energy in cells. Mapping the stress fields in a heterogeneous time-dependent environment like that found in cells requires probes that are specific for different proteins and respond to biologically relevant forces with minimal disturbance to the host system. To meet these goals, we have designed a genetically encoded stress sensor with minimal volume and high sensitivity and dynamic range. The new FRET-based sensor, called cpstFRET, is designed to be modulated by the angles between the donor and acceptor rather than the distance between them. Relative to other probes, it is physically smaller and exhibits a greater dynamic range and sensitivity and expresses well. For in vivo testing, we measured stress gradients in time and space in non-erythroid spectrin in several different cell types and found that spectrin is under constitutive stress in some cells but not in others. Stresses appear to be generated by both F-actin and tubulin. The probe revealed, for the first time, that spectrin undergoes time-dependent force modulation during cell migration. cpstFRET can be employed in vitro, in vivo and in situ, and when incorporated into biologically expressed extracellular polymers such as collagen, it can report multidimensional stress fields.
Fanjie Meng; Frederick Sachs
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of cell science     Volume:  125     ISSN:  1477-9137     ISO Abbreviation:  J. Cell. Sci.     Publication Date:  2012 Feb 
Date Detail:
Created Date:  2012-03-05     Completed Date:  2012-09-06     Revised Date:  2014-09-18    
Medline Journal Info:
Nlm Unique ID:  0052457     Medline TA:  J Cell Sci     Country:  England    
Other Details:
Languages:  eng     Pagination:  743-50     Citation Subset:  IM    
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MeSH Terms
Actins / metabolism
Bacterial Proteins / chemistry,  genetics
Base Sequence
Biomechanical Phenomena
Biophysical Phenomena
Cell Line
Cell Movement / physiology*
Computer Systems
Cytoskeleton / metabolism
DNA Primers / genetics
Fluorescence Resonance Energy Transfer / methods*
Fluorescent Dyes / chemistry
HEK293 Cells
Luminescent Proteins / chemistry,  genetics
Models, Molecular
Protein Multimerization
Recombinant Proteins / chemistry,  genetics
Spectrin / chemistry,  metabolism*
Stress, Mechanical
Tubulin / metabolism
Grant Support
Reg. No./Substance:
0/Actins; 0/Bacterial Proteins; 0/DNA Primers; 0/Fluorescent Dyes; 0/Luminescent Proteins; 0/Recombinant Proteins; 0/Tubulin; 0/yellow fluorescent protein, Bacteria; 12634-43-4/Spectrin

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