title: Mechanically Induced Catalytic Amplification Reaction for Readout of Receptor-Mediated Cellular Forces.
abstract: Mechanics play a fundamental role in cell biology, but detecting piconewton (pN) forces is challenging because of a lack of accessible and high throughput assays. A mechanically induced catalytic amplification reaction (MCR) for readout of receptor-mediated forces in cells is described. Mechanically labile DNA duplexes presenting ligands are surface immobilized such that specific receptor forces denature the duplex and thus expose a blocked primer. Amplification of primers is achieved using an isothermal polymerization reaction and quantified by fluorescence readout. As a proof of concept, the assay was used to test the activity of a mechanomodulatory drug and integrin adhesion receptor antibodies. To the best of our knowledge, this is the first example of a catalytic reaction triggered in response to molecular piconewton forces. The MCR may transform the field of mechanobiology by providing a new facile tool to detect receptor specific mechanics with the convenience of the polymerase chain reaction (PCR).
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA, 30322, USA.
authors.forename: Victor Pui-Yan
authors.initials: VP
authors.lastname: Ma
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA, 30322, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA, 30322, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Yehl
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA, 30322, USA.
authors.forename: Kornelia
authors.initials: K
authors.lastname: Galior
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA, 30322, USA.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Chemistry, Emory University, Atlanta, GA, 30322, USA. k.salaita@emory.edu.|Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, 30322, USA. k.salaita@emory.edu.
authors.forename: Khalid
authors.initials: K
authors.lastname: Salaita
mesh_terms: D000818:Animals; D055162:Biocatalysis; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D015394:Molecular Structure; D041681:NIH 3T3 Cells; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA; D011088:DNA Ligases
keywords: drug screening; integrins; mechanically induced catalytic amplification; receptor-mediated forces; rolling circle amplification