Computational and experimental approaches to quantify protein binding interactions under confinement.

Autor:	Leckband D; Department of Chemical and Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois; Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois; Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois. Electronic address: leckband@illinois.edu., Schwartz DK; Chemical and Biomolecular Engineering, University of Colorado Boulder, Boulder, Colorado., Wu Y; Department of Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, New York.
Jazyk:	angličtina
Zdroj:	Biophysical journal [Biophys J] 2024 Feb 20; Vol. 123 (4), pp. 424-434. Date of Electronic Publication: 2024 Jan 20.
DOI:	10.1016/j.bpj.2024.01.018
Abstrakt:	Crowded environments and confinement alter the interactions of adhesion proteins confined to membranes or narrow, crowded gaps at adhesive contacts. Experimental approaches and theoretical frameworks were developed to quantify protein binding constants in these environments. However, recent predictions and the complexity of some protein interactions proved challenging to address with prior experimental or theoretical approaches. This perspective highlights new methods developed by these authors that address these challenges. Specifically, single-molecule fluorescence resonance energy transfer and single-molecule tracking measurements were developed to directly image the binding/unbinding rates of membrane-tethered cadherins. Results identified predicted cis (lateral) interactions, which control cadherin clustering on membranes but were not detected in solution. Kinetic Monte Carlo simulations, based on a realistic model of cis cadherin interactions, were developed to extract binding/unbinding rate constants from heterogeneous single-molecule data. The extension of single-molecule fluorescence measurements to cis and trans (adhesive) cadherin interactions at membrane junctions identified unexpected cooperativity between cis and trans binding that appears to enhance intercellular binding kinetics. Comparisons of intercellular binding kinetics, kinetic Monte Carlo simulations, and single-molecule fluorescence data suggest a strategy to bridge protein binding kinetics across length scales. Although cadherin is the focus of these studies, the approaches can be extended to other intercellular adhesion proteins. Competing Interests: Declaration of interests The authors declare that they have no competing interests. (Copyright © 2024 Biophysical Society. Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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