Skip to main content

Cryo-EM and molecular docking shows myosin-S1 loop 4 contacts actin and tropomyosin on thin filaments

Doran, Matthew H., Pavadai, Elumalai, Rynkiewicz, Michael J., Walklate, Jonathan, Bullitt, Esther, Moore, Jeffrey R., Regnier, Michael, Geeves, Michael A., Lehman, William (2020) Cryo-EM and molecular docking shows myosin-S1 loop 4 contacts actin and tropomyosin on thin filaments. Biophysical Journal, . ISSN 0006-3495. (In press) (doi:10.1016/j.bpj.2020.07.006) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:82133)

PDF Author's Accepted Manuscript
Language: English

Restricted to Repository staff only
Contact us about this Publication
Official URL
https://doi.org/10.1016/j.bpj.2020.07.006

Abstract

The motor protein, myosin, drives muscle and non-muscle motility by binding to and moving along actin of thin filaments. Myosin-binding to actin also modulates interactions of the regulatory protein, tropomyosin, on thin filaments, and conversely tropomyosin affects myosin-binding to actin. Insight into this reciprocity will facilitate a molecular level elucidation of tropomyosin regulation of myosin interaction with actin in muscle contraction, and in turn, promote better understanding non-muscle cell motility. Indeed, experimental approaches, such as fiber diffraction, cryo-electron microscopy and 3D reconstruction, have long been used to define regulatory interaction of tropomyosin and myosin on actin at a structural level. However, their limited resolution has not proven sufficient to determine tropomyosin and myosin contacts at an atomic-level and thus to fully substantiate possible functional contributions. To overcome this deficiency, we have followed a hybrid approach by performing new cryo-EM reconstruction of myosin-S1‒decorated F-actin-tropomyosin together with atomic-scale protein-protein docking of tropomyosin to the EM models. Here, cryo-EM data were derived from filaments reconstituted with α1-actin, cardiac αα-tropomyosin, and masseter muscle β-myosin complexes; masseter myosin, which shares sequence identity with β-cardiac myosin-heavy chain, was used because of its stability in vitro. The data were used to build an atomic model of the tropomyosin cable that fits onto the actin filament between the tip of the myosin head and a cleft on the innermost edge of actin subunits. The docking and atomic scale fitting showed multiple discrete interactions of myosin loop 4 and acidic residues on successive 39 to 42 residue-long tropomyosin pseudo-repeats. The contacts between S1 and tropomyosin on actin appear to compete with and displace ones normally found between actin and tropomyosin on myosin-free thin filaments in relaxed muscle, thus restructuring the filament during myosin-induced activation.

Item Type: Article
DOI/Identification number: 10.1016/j.bpj.2020.07.006
Uncontrolled keywords: Actin, ClusPro, molecular dynamics, myosin, tropomyosin
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Susan Davies
Date Deposited: 16 Jul 2020 15:01 UTC
Last Modified: 16 Jul 2020 15:02 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/82133 (The current URI for this page, for reference purposes)
Geeves, Michael A.: https://orcid.org/0000-0002-9364-8898
  • Depositors only (login required):