Integrin-generated forces lead to streptavidin-biotin unbinding in cellular adhesions.
The interplay between chemical and mechanical signals plays an important role in cell biology, and integrin receptors are the primary molecules involved in sensing and transducing external mechanical cues. We used integrin-specific probes in molecular tension fluorescence microscopy to investigate the pN forces exerted by integrin receptors in living cells. The molecular tension fluorescence microscopy probe consisted of a cyclic Arg-Gly-Asp-D-Phe-Lys(Cys) (cRGDfK(C)) peptide tethered to the terminus of a polyethylene glycol polymer that was attached to a surface through streptavidin-biotin linkage. A fluorescence resonance energy transfer mechanism was used to visualize tension-driven extension of the polymer. Surprisingly, we found that integrin receptors dissociate streptavidin-biotin tethered ligands in focal adhesions within 60 min of cell seeding. Although streptavidin-biotin binding affinity is described as the strongest noncovalent bond in nature, and is ~10(6) - 10(8) times larger than that of integrin-RGD affinity, our results suggest that individual integrin-ligand complexes undergo a marked enhancement in stability when the receptor assembles in the cell membrane. Based on the observation of streptavidin-biotin unbinding, we also conclude that the magnitude of integrin-ligand tension in focal adhesions can reach values that are at least 10 fold larger than was previously estimated using traction force microscopy-based methods.
Department of Chemistry, Emory University, Atlanta, Georgia.
Carol
C
Jurchenko
Department of Chemistry, Emory University, Atlanta, Georgia.
Yuan
Y
Chang
Department of Chemistry, Emory University, Atlanta, Georgia.
Yoshie
Y
Narui
Department of Chemistry, Emory University, Atlanta, Georgia.
Yun
Y
Zhang
Department of Chemistry, Emory University, Atlanta, Georgia. Electronic address: k.salaita@emory.edu.
Khalid S
KS
Salaita
D000818:Animals; D001710:Biotin / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology*; D002452:Cell Count; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D055595:Mechanical Phenomena; D051379:Mice; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D010455:Peptides / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D019809:Streptavidin / Q000378:metabolism*; D013997:Time Factors
D016023:Integrins; D010455:Peptides; D011092:Polyethylene Glycols; D001710:Biotin; D019809:Streptavidin