Titin-actin interaction in mouse myocardium: Passive tension modulation and its regulation by calcium/S100A1

R. Yamasaki, M. Berri, Y. Wu, K. Trombitás, M. McNabb, M. S Z Kellermayer, C. Witt, D. Labeit, S. Labeit, M. Greaser, Hendrikus "Henk" Granzier

Research output: Contribution to journalArticle

170 Citations (Scopus)

Abstract

Passive tension in striated muscles derives primarily from the extension of the giant protein titin. However, several studies have suggested that, in cardiac muscle, interactions between titin and actin might also contribute to passive tension. We expressed recombinant fragments representing the subdomains of the extensible region of cardiac N2B titin (tandem-Ig segments, the N2B splice element, and the PEVK domain), and assayed them for binding to F-actin. The PEVK fragment bound F-actin, but no binding was detected for the other fragments. Comparison with a skeletal muscle PEVK fragment revealed that only the cardiac PEVK binds actin at physiological ionic strengths. The significance of PEVK-actin interaction was investigated using in vitro motility and single-myocyte mechanics. As F-actin slid relative to titin in the motility assay, a dynamic interaction between the PEVK domain and F-actin retarded filament sliding. Myocyte results suggest that a similar interaction makes a significant contribution to the passive tension. We also investigated the effect of calcium on PEVK-actin interaction. Although calcium alone had no effect, S100A1, a soluble calcium-binding protein found at high concentrations in the myocardium, inhibited PEVK-actin interaction in a calcium-dependent manner. Gel overlay analysis revealed that S100A1 bound the PEVK region in vitro in a calcium-dependent manner, and S100A1 binding was observed at several sites along titin's extensible region in situ, including the PEVK domain. In vitro motility results indicate that S100A1-PEVK interaction reduces the force that arises as F-actin slides relative to the PEVK domain, and we speculate that S100A1 may provide a mechanism to free the thin filament from titin and reduce titin-based tension before active contraction.

Original languageEnglish (US)
Pages (from-to)2297-2313
Number of pages17
JournalBiophysical Journal
Volume81
Issue number4
StatePublished - 2001
Externally publishedYes

Fingerprint

Connectin
Actins
Myocardium
Calcium
Muscle Cells
Calcium-Binding Proteins
Striated Muscle
Mechanics
Actin Cytoskeleton
Osmolar Concentration
Skeletal Muscle
Gels

ASJC Scopus subject areas

  • Biophysics

Cite this

Yamasaki, R., Berri, M., Wu, Y., Trombitás, K., McNabb, M., Kellermayer, M. S. Z., ... Granzier, H. H. (2001). Titin-actin interaction in mouse myocardium: Passive tension modulation and its regulation by calcium/S100A1. Biophysical Journal, 81(4), 2297-2313.

Titin-actin interaction in mouse myocardium : Passive tension modulation and its regulation by calcium/S100A1. / Yamasaki, R.; Berri, M.; Wu, Y.; Trombitás, K.; McNabb, M.; Kellermayer, M. S Z; Witt, C.; Labeit, D.; Labeit, S.; Greaser, M.; Granzier, Hendrikus "Henk".

In: Biophysical Journal, Vol. 81, No. 4, 2001, p. 2297-2313.

Research output: Contribution to journalArticle

Yamasaki, R, Berri, M, Wu, Y, Trombitás, K, McNabb, M, Kellermayer, MSZ, Witt, C, Labeit, D, Labeit, S, Greaser, M & Granzier, HH 2001, 'Titin-actin interaction in mouse myocardium: Passive tension modulation and its regulation by calcium/S100A1', Biophysical Journal, vol. 81, no. 4, pp. 2297-2313.
Yamasaki R, Berri M, Wu Y, Trombitás K, McNabb M, Kellermayer MSZ et al. Titin-actin interaction in mouse myocardium: Passive tension modulation and its regulation by calcium/S100A1. Biophysical Journal. 2001;81(4):2297-2313.
Yamasaki, R. ; Berri, M. ; Wu, Y. ; Trombitás, K. ; McNabb, M. ; Kellermayer, M. S Z ; Witt, C. ; Labeit, D. ; Labeit, S. ; Greaser, M. ; Granzier, Hendrikus "Henk". / Titin-actin interaction in mouse myocardium : Passive tension modulation and its regulation by calcium/S100A1. In: Biophysical Journal. 2001 ; Vol. 81, No. 4. pp. 2297-2313.
@article{9f335e3497344f8cba51b1c129fa6bcd,
title = "Titin-actin interaction in mouse myocardium: Passive tension modulation and its regulation by calcium/S100A1",
abstract = "Passive tension in striated muscles derives primarily from the extension of the giant protein titin. However, several studies have suggested that, in cardiac muscle, interactions between titin and actin might also contribute to passive tension. We expressed recombinant fragments representing the subdomains of the extensible region of cardiac N2B titin (tandem-Ig segments, the N2B splice element, and the PEVK domain), and assayed them for binding to F-actin. The PEVK fragment bound F-actin, but no binding was detected for the other fragments. Comparison with a skeletal muscle PEVK fragment revealed that only the cardiac PEVK binds actin at physiological ionic strengths. The significance of PEVK-actin interaction was investigated using in vitro motility and single-myocyte mechanics. As F-actin slid relative to titin in the motility assay, a dynamic interaction between the PEVK domain and F-actin retarded filament sliding. Myocyte results suggest that a similar interaction makes a significant contribution to the passive tension. We also investigated the effect of calcium on PEVK-actin interaction. Although calcium alone had no effect, S100A1, a soluble calcium-binding protein found at high concentrations in the myocardium, inhibited PEVK-actin interaction in a calcium-dependent manner. Gel overlay analysis revealed that S100A1 bound the PEVK region in vitro in a calcium-dependent manner, and S100A1 binding was observed at several sites along titin's extensible region in situ, including the PEVK domain. In vitro motility results indicate that S100A1-PEVK interaction reduces the force that arises as F-actin slides relative to the PEVK domain, and we speculate that S100A1 may provide a mechanism to free the thin filament from titin and reduce titin-based tension before active contraction.",
author = "R. Yamasaki and M. Berri and Y. Wu and K. Trombit{\'a}s and M. McNabb and Kellermayer, {M. S Z} and C. Witt and D. Labeit and S. Labeit and M. Greaser and Granzier, {Hendrikus {"}Henk{"}}",
year = "2001",
language = "English (US)",
volume = "81",
pages = "2297--2313",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "4",

}

TY - JOUR

T1 - Titin-actin interaction in mouse myocardium

T2 - Passive tension modulation and its regulation by calcium/S100A1

AU - Yamasaki, R.

AU - Berri, M.

AU - Wu, Y.

AU - Trombitás, K.

AU - McNabb, M.

AU - Kellermayer, M. S Z

AU - Witt, C.

AU - Labeit, D.

AU - Labeit, S.

AU - Greaser, M.

AU - Granzier, Hendrikus "Henk"

PY - 2001

Y1 - 2001

N2 - Passive tension in striated muscles derives primarily from the extension of the giant protein titin. However, several studies have suggested that, in cardiac muscle, interactions between titin and actin might also contribute to passive tension. We expressed recombinant fragments representing the subdomains of the extensible region of cardiac N2B titin (tandem-Ig segments, the N2B splice element, and the PEVK domain), and assayed them for binding to F-actin. The PEVK fragment bound F-actin, but no binding was detected for the other fragments. Comparison with a skeletal muscle PEVK fragment revealed that only the cardiac PEVK binds actin at physiological ionic strengths. The significance of PEVK-actin interaction was investigated using in vitro motility and single-myocyte mechanics. As F-actin slid relative to titin in the motility assay, a dynamic interaction between the PEVK domain and F-actin retarded filament sliding. Myocyte results suggest that a similar interaction makes a significant contribution to the passive tension. We also investigated the effect of calcium on PEVK-actin interaction. Although calcium alone had no effect, S100A1, a soluble calcium-binding protein found at high concentrations in the myocardium, inhibited PEVK-actin interaction in a calcium-dependent manner. Gel overlay analysis revealed that S100A1 bound the PEVK region in vitro in a calcium-dependent manner, and S100A1 binding was observed at several sites along titin's extensible region in situ, including the PEVK domain. In vitro motility results indicate that S100A1-PEVK interaction reduces the force that arises as F-actin slides relative to the PEVK domain, and we speculate that S100A1 may provide a mechanism to free the thin filament from titin and reduce titin-based tension before active contraction.

AB - Passive tension in striated muscles derives primarily from the extension of the giant protein titin. However, several studies have suggested that, in cardiac muscle, interactions between titin and actin might also contribute to passive tension. We expressed recombinant fragments representing the subdomains of the extensible region of cardiac N2B titin (tandem-Ig segments, the N2B splice element, and the PEVK domain), and assayed them for binding to F-actin. The PEVK fragment bound F-actin, but no binding was detected for the other fragments. Comparison with a skeletal muscle PEVK fragment revealed that only the cardiac PEVK binds actin at physiological ionic strengths. The significance of PEVK-actin interaction was investigated using in vitro motility and single-myocyte mechanics. As F-actin slid relative to titin in the motility assay, a dynamic interaction between the PEVK domain and F-actin retarded filament sliding. Myocyte results suggest that a similar interaction makes a significant contribution to the passive tension. We also investigated the effect of calcium on PEVK-actin interaction. Although calcium alone had no effect, S100A1, a soluble calcium-binding protein found at high concentrations in the myocardium, inhibited PEVK-actin interaction in a calcium-dependent manner. Gel overlay analysis revealed that S100A1 bound the PEVK region in vitro in a calcium-dependent manner, and S100A1 binding was observed at several sites along titin's extensible region in situ, including the PEVK domain. In vitro motility results indicate that S100A1-PEVK interaction reduces the force that arises as F-actin slides relative to the PEVK domain, and we speculate that S100A1 may provide a mechanism to free the thin filament from titin and reduce titin-based tension before active contraction.

UR - http://www.scopus.com/inward/record.url?scp=0034811015&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034811015&partnerID=8YFLogxK

M3 - Article

C2 - 11566799

AN - SCOPUS:0034811015

VL - 81

SP - 2297

EP - 2313

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 4

ER -