title: Spider Silk Peptide Is a Compact, Linear Nanospring Ideal for Intracellular Tension Sensing.
abstract: Recent development and applications of calibrated, fluorescence resonance energy transfer (FRET)-based tension sensors have led to a new understanding of single molecule mechanotransduction in a number of biological systems. To expand the range of accessible forces, we systematically measured FRET versus force trajectories for 25, 40, and 50 amino acid peptide repeats derived from spider silk. Single molecule fluorescence-force spectroscopy showed that the peptides behaved as linear springs instead of the nonlinear behavior expected for a disordered polymer. Our data are consistent with a compact, rodlike structure that measures 0.26 nm per 5 amino acid repeat that can stretch by 500% while maintaining linearity, suggesting that the remarkable elasticity of spider silk proteins may in part derive from the properties of individual chains. We found the shortest peptide to have the widest range of force sensitivity: between 2 pN and 11 pN. Live cell imaging of the three tension sensor constructs inserted into vinculin showed similar force values around 2.4 pN. We also provide a lookup table for force versus intracellular FRET for all three constructs.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Brenner
authors.forename: Ruobo
authors.initials: R
authors.lastname: Zhou
authors.affiliation: Cardiovascular Research Center, University of Virginia , Charlottesville, Virginia 22908, United States.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Conway
authors.affiliation: Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California , Irvine, California 92697, United States.
authors.forename: Luca
authors.initials: L
authors.lastname: Lanzano
authors.affiliation: Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California , Irvine, California 92697, United States.
authors.forename: Enrico
authors.initials: E
authors.lastname: Gratton
authors.affiliation: Cardiovascular Research Center, University of Virginia , Charlottesville, Virginia 22908, United States.|Yale Cardiovascular Research Center, Departments of Internal Medicine (Section of Cardiovascular Medicine) and Cell Biology, Yale University , New Haven, Connecticut 06511, United States.
authors.forename: Martin A
authors.initials: MA
authors.lastname: Schwartz
authors.affiliation: Institute for Genomic Biology, University of Illinois at Urbana-Champaign , Urbana, Illinois 61801, United States.|Department of Biophysics & Biophysical Chemistry, Department of Biophysics, and Department of Biomedical Engineering, Johns Hopkins University , Baltimore, Maryland 21205, United States.|Howard Hughes Medical Institute , Baltimore, Maryland 21205, United States.
authors.forename: Taekjip
authors.initials: T
authors.lastname: Ha
mesh_terms: D000818:Animals; D004548:Elasticity; D031541:Fluorescence Resonance Energy Transfer; D040542:Mechanotransduction, Cellular; D010455:Peptides / Q000737:chemistry*; D047011:Silk / Q000737:chemistry*; D013112:Spiders / Q000737:chemistry*; D013314:Stress, Mechanical
chemical_list: D010455:Peptides; D047011:Silk
keywords: FLIM; Force-sensor; force-fluorescence spectroscopy; optical tweezers; single-molecule FRET; spider flagelliform silk
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