西亚试剂：Nanoscale architecture of integrin-based cell adhesions

Nanoscale architecture of integrin-based cell adhesions
Pakorn Kanchanawong1,5, Gleb Shtengel2,5, Ana M. Pasapera1, Ericka B. Ramko3, Michael W. Davidson3,4, Harald F. Hess2 & Clare M. Waterman1

1.National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
2.Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia 20147, USA
3.National High Magnetic Field Laboratory, The Florida State University, Tallahassee, Florida 32310, USA
4.Department of Biological Science, The Florida State University, Tallahassee, Florida 32306, USA
5.These authors contributed equally to this work.
Correspondence to: Clare M. Waterman1 Email: watermancm@nhlbi.nih.gov

Correspondence to: Harald F. Hess2 Email: hessh@janelia.hhmi.org

Correspondence to: Michael W. Davidson3,4 Email: davidson@magnet.fsu.edu

Top of pageAbstractCell adhesions to the extracellular matrix (ECM) are necessary for morphogenesis, immunity and wound healing1, 2. Focal adhesions are multifunctional organelles that mediate cell–ECM adhesion, force transmission, cytoskeletal regulation and signalling1, 2, 3. Focal adhesions consist of a complex network4 of trans-plasma-membrane integrins and cytoplasmic proteins that form a?<200-nm plaque5, 6 linking the ECM to the actin cytoskeleton. The complexity of focal adhesion composition and dynamics implicate an intricate molecular machine7, 8. However, focal adhesion molecular architecture remains unknown. Here we used three-dimensional super-resolution fluorescence microscopy (interferometric photoactivated localization microscopy)9 to map nanoscale protein organization in focal adhesions. Our results reveal that integrins and actin are vertically separated by a ~40-nm focal adhesion core region consisting of multiple protein-specific strata: a membrane-apposed integrin signalling layer containing integrin cytoplasmic tails, focal adhesion kinase and paxillin; an intermediate force-transduction layer containing talin and vinculin; and an uppermost actin-regulatory layer containing zyxin, vasodilator-stimulated phosphoprotein and α-actinin. By localizing amino- and carboxy-terminally tagged talins, we reveal talin’s polarized orientation, indicative of a role in organizing the focal adhesion strata. The composite multilaminar protein architecture provides a molecular blueprint for understanding focal adhesion functions.