Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments

Tropomodulin (Tmod) is an actin-binding protein that interacts with tropomyosin and the actin filament at the pointed end. The influence of Tmod on the thin filament activation in the myocardium is not clear. We studied the interactions of Tmod1 and Tmod4 with the cardiac tropomyosin isoforms Tpm1.1...

Full description

Saved in:
Bibliographic Details
Main Authors: Galina V. Kopylova, Anastasia M. Kochurova, Evgeniia A. Beldiia, Andrey V. Slushchev, Victoria V. Nefedova, Natalia S. Ryabkova, Ivan A. Katrukha, Daria S. Yampolskaya, Alexander M. Matyushenko, Daniil V. Shchepkin
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Biomolecules
Subjects:
Online Access:https://www.mdpi.com/2218-273X/15/5/727
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850127615929614336
author Galina V. Kopylova
Anastasia M. Kochurova
Evgeniia A. Beldiia
Andrey V. Slushchev
Victoria V. Nefedova
Natalia S. Ryabkova
Ivan A. Katrukha
Daria S. Yampolskaya
Alexander M. Matyushenko
Daniil V. Shchepkin
author_facet Galina V. Kopylova
Anastasia M. Kochurova
Evgeniia A. Beldiia
Andrey V. Slushchev
Victoria V. Nefedova
Natalia S. Ryabkova
Ivan A. Katrukha
Daria S. Yampolskaya
Alexander M. Matyushenko
Daniil V. Shchepkin
author_sort Galina V. Kopylova
collection DOAJ
description Tropomodulin (Tmod) is an actin-binding protein that interacts with tropomyosin and the actin filament at the pointed end. The influence of Tmod on the thin filament activation in the myocardium is not clear. We studied the interactions of Tmod1 and Tmod4 with the cardiac tropomyosin isoforms Tpm1.1 and Tpm1.2 using size-exclusion chromatography, a pull-down assay, and cross-linking with glutaraldehyde. We found that Tmod1 and Tmod4 form complexes with both Tpm1.1 and Tpm1.2, indicating durable interactions between these proteins. The effects of both Tmods on the actin–myosin interaction were studied using an in vitro motility assay. Tmod did not affect the sliding velocity of bare F-actin. Tmod1 slightly dose-dependently decreased the sliding velocity of F-actin–Tpm1.1 filaments and had no effect on the velocity of F-actin–Tpm1.2 filaments. With ventricular myosin, Tmod1 reduced the calcium sensitivity of the sliding velocity of thin filaments containing Tpm1.1 but did not affect it with filaments containing Tpm1.2. With atrial myosin, Tmod1 decreased the calcium sensitivity of the sliding velocities of thin filaments containing both Tpm1.1 and Tpm1.2. We can conclude that Tmod takes part in the regulation of actin–myosin interactions in the myocardium through interactions with Tpm. The effect of Tmod on the activation of thin filaments depends on the protein isoforms.
format Article
id doaj-art-5fc364c181c44a1ca3a0237d99263252
institution OA Journals
issn 2218-273X
language English
publishDate 2025-05-01
publisher MDPI AG
record_format Article
series Biomolecules
spelling doaj-art-5fc364c181c44a1ca3a0237d992632522025-08-20T02:33:38ZengMDPI AGBiomolecules2218-273X2025-05-0115572710.3390/biom15050727Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin FilamentsGalina V. Kopylova0Anastasia M. Kochurova1Evgeniia A. Beldiia2Andrey V. Slushchev3Victoria V. Nefedova4Natalia S. Ryabkova5Ivan A. Katrukha6Daria S. Yampolskaya7Alexander M. Matyushenko8Daniil V. Shchepkin9Institute of Immunology and Physiology of the Russian Academy of Sciences, Yekaterinburg 620049, RussiaInstitute of Immunology and Physiology of the Russian Academy of Sciences, Yekaterinburg 620049, RussiaInstitute of Immunology and Physiology of the Russian Academy of Sciences, Yekaterinburg 620049, RussiaResearch Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, RussiaResearch Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, RussiaDepartment of Biochemistry, Faculty of Biology, Lomonosov Moscow State University, Moscow 119991, RussiaDepartment of Biochemistry, Faculty of Biology, Lomonosov Moscow State University, Moscow 119991, RussiaResearch Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, RussiaResearch Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, RussiaInstitute of Immunology and Physiology of the Russian Academy of Sciences, Yekaterinburg 620049, RussiaTropomodulin (Tmod) is an actin-binding protein that interacts with tropomyosin and the actin filament at the pointed end. The influence of Tmod on the thin filament activation in the myocardium is not clear. We studied the interactions of Tmod1 and Tmod4 with the cardiac tropomyosin isoforms Tpm1.1 and Tpm1.2 using size-exclusion chromatography, a pull-down assay, and cross-linking with glutaraldehyde. We found that Tmod1 and Tmod4 form complexes with both Tpm1.1 and Tpm1.2, indicating durable interactions between these proteins. The effects of both Tmods on the actin–myosin interaction were studied using an in vitro motility assay. Tmod did not affect the sliding velocity of bare F-actin. Tmod1 slightly dose-dependently decreased the sliding velocity of F-actin–Tpm1.1 filaments and had no effect on the velocity of F-actin–Tpm1.2 filaments. With ventricular myosin, Tmod1 reduced the calcium sensitivity of the sliding velocity of thin filaments containing Tpm1.1 but did not affect it with filaments containing Tpm1.2. With atrial myosin, Tmod1 decreased the calcium sensitivity of the sliding velocities of thin filaments containing both Tpm1.1 and Tpm1.2. We can conclude that Tmod takes part in the regulation of actin–myosin interactions in the myocardium through interactions with Tpm. The effect of Tmod on the activation of thin filaments depends on the protein isoforms.https://www.mdpi.com/2218-273X/15/5/727tropomodulincardiac tropomyosin isoformsactin-associated proteinscardiac myosin isoformsactin–myosin interactioncalcium regulation
spellingShingle Galina V. Kopylova
Anastasia M. Kochurova
Evgeniia A. Beldiia
Andrey V. Slushchev
Victoria V. Nefedova
Natalia S. Ryabkova
Ivan A. Katrukha
Daria S. Yampolskaya
Alexander M. Matyushenko
Daniil V. Shchepkin
Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments
Biomolecules
tropomodulin
cardiac tropomyosin isoforms
actin-associated proteins
cardiac myosin isoforms
actin–myosin interaction
calcium regulation
title Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments
title_full Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments
title_fullStr Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments
title_full_unstemmed Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments
title_short Tropomodulin–Tropomyosin Interplay Modulates Interaction Between Cardiac Myosin and Thin Filaments
title_sort tropomodulin tropomyosin interplay modulates interaction between cardiac myosin and thin filaments
topic tropomodulin
cardiac tropomyosin isoforms
actin-associated proteins
cardiac myosin isoforms
actin–myosin interaction
calcium regulation
url https://www.mdpi.com/2218-273X/15/5/727
work_keys_str_mv AT galinavkopylova tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT anastasiamkochurova tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT evgeniiaabeldiia tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT andreyvslushchev tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT victoriavnefedova tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT nataliasryabkova tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT ivanakatrukha tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT dariasyampolskaya tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT alexandermmatyushenko tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments
AT daniilvshchepkin tropomodulintropomyosininterplaymodulatesinteractionbetweencardiacmyosinandthinfilaments