Publication Date:
2010-07-09
Description:
Mechanical forces are central to developmental, physiological and pathological processes. However, limited understanding of force transmission within sub-cellular structures is a major obstacle to unravelling molecular mechanisms. Here we describe the development of a calibrated biosensor that measures forces across specific proteins in cells with piconewton (pN) sensitivity, as demonstrated by single molecule fluorescence force spectroscopy. The method is applied to vinculin, a protein that connects integrins to actin filaments and whose recruitment to focal adhesions (FAs) is force-dependent. We show that tension across vinculin in stable FAs is approximately 2.5 pN and that vinculin recruitment to FAs and force transmission across vinculin are regulated separately. Highest tension across vinculin is associated with adhesion assembly and enlargement. Conversely, vinculin is under low force in disassembling or sliding FAs at the trailing edge of migrating cells. Furthermore, vinculin is required for stabilizing adhesions under force. Together, these data reveal that FA stabilization under force requires both vinculin recruitment and force transmission, and that, surprisingly, these processes can be controlled independently.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2901888/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2901888/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Grashoff, Carsten -- Hoffman, Brenton D -- Brenner, Michael D -- Zhou, Ruobo -- Parsons, Maddy -- Yang, Michael T -- McLean, Mark A -- Sligar, Stephen G -- Chen, Christopher S -- Ha, Taekjip -- Schwartz, Martin A -- 5T32-HL007284/HL/NHLBI NIH HHS/ -- R01 GM033775/GM/NIGMS NIH HHS/ -- R21 RR025341/RR/NCRR NIH HHS/ -- U54 GM064346/GM/NIGMS NIH HHS/ -- U54 GM064346-099039/GM/NIGMS NIH HHS/ -- U54 GM64346/GM/NIGMS NIH HHS/ -- Howard Hughes Medical Institute/ -- England -- Nature. 2010 Jul 8;466(7303):263-6. doi: 10.1038/nature09198.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Robert M. Berne Cardiovascular Research Center, University of Virginia, Charlottesville, Virginia 22908, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/20613844" target="_blank"〉PubMed〈/a〉
Keywords:
Animals
;
Biosensing Techniques
;
Calibration
;
Cattle
;
Cell Line
;
Cell Movement/*physiology
;
Fluorescent Dyes
;
Focal Adhesions/*metabolism
;
Humans
;
Mice
;
Microscopy, Confocal
;
Movement
;
Optical Tweezers
;
Spectrometry, Fluorescence
;
*Stress, Mechanical
;
Vinculin/chemistry/deficiency/genetics/*metabolism
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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