Constructing modular and universal single molecule tension sensor using protein G to study mechano-sensitive receptors

Xuefeng Wang, Zainab Rahil, Isaac T.S. Li, Farhan Chowdhury, Deborah E. Leckband, Yann R. Chemla, Taekjip Ha

Research output: Contribution to journalArticle

Abstract

Recently a variety of molecular force sensors have been developed to study cellular forces acting through single mechano-sensitive receptors. A common strategy adopted is to attach ligand molecules on a surface through engineered molecular tethers which report cell-exerted tension on receptor-ligand bonds. This approach generally requires chemical conjugation of the ligand to the force reporting tether which can be time-consuming and labor-intensive. Moreover, ligand-tether conjugation can severely reduce the activity of protein ligands. To address this problem, we developed a Protein G (ProG)-based force sensor in which force-reporting tethers are conjugated to ProG instead of ligands. A recombinant ligand fused with IgG-Fc is conveniently assembled with the force sensor through ProG:Fc binding, therefore avoiding ligand conjugation and purification processes. Using this approach, we determined that molecular tension on E-cadherin is lower than dsDNA unzipping force (nominal value: 12 pN) during initial cadherin-mediated cell adhesion, followed by an escalation to forces higher than 43 pN (nominal value). This approach is highly modular and potentially universal as we demonstrate using two additional receptor-ligand interactions, P-selectin & PSGL-1 and Notch & DLL1.

Original languageEnglish (US)
Article number21584
JournalScientific reports
Volume6
DOIs
StatePublished - Feb 15 2016

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Ligands
Proteins
Cadherins
P-Selectin
Gravitation
Cell Adhesion
Immunoglobulin G

ASJC Scopus subject areas

  • General

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Constructing modular and universal single molecule tension sensor using protein G to study mechano-sensitive receptors. / Wang, Xuefeng; Rahil, Zainab; Li, Isaac T.S.; Chowdhury, Farhan; Leckband, Deborah E.; Chemla, Yann R.; Ha, Taekjip.

In: Scientific reports, Vol. 6, 21584, 15.02.2016.

Research output: Contribution to journalArticle

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