TY - JOUR

T1 - Probing Ligand-Receptor Interaction in Living Cells Using Force Measurements With Optical Tweezers

AU - Riesenberg, Carolin

AU - Iriarte-Valdez, Christian Alejandro

AU - Becker, Annegret

AU - Dienerowitz, Maria

AU - Heisterkamp, Alexander

AU - Ngezahayo, Anaclet

AU - Torres, Maria Leilani

N1 - Funding Information: This work was supported by the German Research Foundation, Germany Clusters of Excellence: REBIRTH (EXC 62) and Hearing4all (EXC 2177). MT-M acknowledges the support of Caroline Herschel Program from the Hochschulbüro für Chancenvielfalt, Leibniz University Hannover.

PY - 2020/11/17

Y1 - 2020/11/17

N2 - This work probes the binding kinetics of COOH-terminus of Clostridium perfringens enterotoxin (c-CPE) and claudin expressing MCF-7 cells using force spectroscopy with optical tweezers. c-CPE is of high biomedical interest due to its ability to specifically bind to claudin with high affinity as well as reversibly disrupt tight junctions whilst maintaining cell viability. We observed single-step rupture events between silica particles functionalized with c-CPE and MCF-7 cells. Extensive calibration of the optical tweezers’ trap stiffness and displacement of the particle from trap center extracted a probable bond rupture force of ≈ 18 pN. The probability of rupture events with c-CPE functionalized silica particles increased by 50% compared to unfunctionalized particles. Additionally, rupture events were not observed when probing cells not expressing claudin with c-CPE coated particles. Overall, this work demonstrates that optical tweezers are invaluable tools to probe ligand-receptor interactions and their potential to study dynamic molecular events in drug-binding scenarios.

AB - This work probes the binding kinetics of COOH-terminus of Clostridium perfringens enterotoxin (c-CPE) and claudin expressing MCF-7 cells using force spectroscopy with optical tweezers. c-CPE is of high biomedical interest due to its ability to specifically bind to claudin with high affinity as well as reversibly disrupt tight junctions whilst maintaining cell viability. We observed single-step rupture events between silica particles functionalized with c-CPE and MCF-7 cells. Extensive calibration of the optical tweezers’ trap stiffness and displacement of the particle from trap center extracted a probable bond rupture force of ≈ 18 pN. The probability of rupture events with c-CPE functionalized silica particles increased by 50% compared to unfunctionalized particles. Additionally, rupture events were not observed when probing cells not expressing claudin with c-CPE coated particles. Overall, this work demonstrates that optical tweezers are invaluable tools to probe ligand-receptor interactions and their potential to study dynamic molecular events in drug-binding scenarios.

KW - C-CPE

KW - cell receptors

KW - claudin

KW - force spectroscopy

KW - ligand

KW - optical tweezers

UR - http://www.scopus.com/inward/record.url?scp=85097076266&partnerID=8YFLogxK

U2 - 10.3389/fbioe.2020.598459

DO - 10.3389/fbioe.2020.598459

M3 - Article

AN - SCOPUS:85097076266

VL - 8

JO - Frontiers in Bioengineering and Biotechnology

JF - Frontiers in Bioengineering and Biotechnology

M1 - 598459

ER -