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Title: Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle

Abstract

Nebulin is a giant sarcomeric protein that spans along the actin filament in skeletal muscle, from the Z-disk to near the thin filament pointed end. Mutations in nebulin cause muscle weakness in nemaline myopathy patients, suggesting that nebulin plays important roles in force generation, yet little is known about nebulin’s influence on thin filament structure and function. Here, we used small-angle X-ray diffraction and compared intact muscle deficient in nebulin (using a conditional nebulin-knockout, Neb cKO) with control (Ctrl) muscle. When muscles were activated, the spacing of the actin subunit repeat (27 Å) increased in both genotypes; when converted to thin filament stiffness, the obtained value was 30 pN/nm in Ctrl muscle and 10 pN/nm in Neb cKO muscle; that is, the thin filament was approximately threefold stiffer when nebulin was present. In contrast, the thick filament stiffness was not different between the genotypes. A significantly shorter left-handed (59 Å) thin filament helical pitch was found in passive and contracting Neb cKO muscles, as well as impaired tropomyosin and troponin movement. Additionally, a reduced myosin mass transfer toward the thin filament in contracting Neb cKO muscle was found, suggesting reduced cross-bridge interaction. Here, we conclude that nebulin is critically importantmore » for physiological force levels, as it greatly stiffens the skeletal muscle thin filament and contributes to thin filament activation and cross-bridge recruitment.« less

Authors:
ORCiD logo [1];  [1];  [2];  [1];  [1];  [2];  [2];  [1]
  1. Univ. of Arizona, Tucson, AZ (United States)
  2. Illinois Inst. of Technology, Chicago, IL (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE; National Institutes of Health (NIH); National Inst. of General Medical Sciences (NIGMS)
OSTI Identifier:
1478092
Grant/Contract Number:  
AC02-06CH11357; P41 GM103622; 1S10OD018090-01
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 115; Journal Issue: 41; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; X-ray diffraction; skeletal myopathy; muscle biology; physiology

Citation Formats

Kiss, Balázs, Lee, Eun-Jeong, Ma, Weikang, Li, Frank W., Tonino, Paola, Mijailovich, Srboljub M., Irving, Thomas C., and Granzier, Henk L. Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle. United States: N. p., 2018. Web. doi:10.1073/pnas.1804726115.
Kiss, Balázs, Lee, Eun-Jeong, Ma, Weikang, Li, Frank W., Tonino, Paola, Mijailovich, Srboljub M., Irving, Thomas C., & Granzier, Henk L. Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle. United States. doi:10.1073/pnas.1804726115.
Kiss, Balázs, Lee, Eun-Jeong, Ma, Weikang, Li, Frank W., Tonino, Paola, Mijailovich, Srboljub M., Irving, Thomas C., and Granzier, Henk L. Mon . "Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle". United States. doi:10.1073/pnas.1804726115. https://www.osti.gov/servlets/purl/1478092.
@article{osti_1478092,
title = {Nebulin stiffens the thin filament and augments cross-bridge interaction in skeletal muscle},
author = {Kiss, Balázs and Lee, Eun-Jeong and Ma, Weikang and Li, Frank W. and Tonino, Paola and Mijailovich, Srboljub M. and Irving, Thomas C. and Granzier, Henk L.},
abstractNote = {Nebulin is a giant sarcomeric protein that spans along the actin filament in skeletal muscle, from the Z-disk to near the thin filament pointed end. Mutations in nebulin cause muscle weakness in nemaline myopathy patients, suggesting that nebulin plays important roles in force generation, yet little is known about nebulin’s influence on thin filament structure and function. Here, we used small-angle X-ray diffraction and compared intact muscle deficient in nebulin (using a conditional nebulin-knockout, Neb cKO) with control (Ctrl) muscle. When muscles were activated, the spacing of the actin subunit repeat (27 Å) increased in both genotypes; when converted to thin filament stiffness, the obtained value was 30 pN/nm in Ctrl muscle and 10 pN/nm in Neb cKO muscle; that is, the thin filament was approximately threefold stiffer when nebulin was present. In contrast, the thick filament stiffness was not different between the genotypes. A significantly shorter left-handed (59 Å) thin filament helical pitch was found in passive and contracting Neb cKO muscles, as well as impaired tropomyosin and troponin movement. Additionally, a reduced myosin mass transfer toward the thin filament in contracting Neb cKO muscle was found, suggesting reduced cross-bridge interaction. Here, we conclude that nebulin is critically important for physiological force levels, as it greatly stiffens the skeletal muscle thin filament and contributes to thin filament activation and cross-bridge recruitment.},
doi = {10.1073/pnas.1804726115},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
issn = {0027-8424},
number = 41,
volume = 115,
place = {United States},
year = {2018},
month = {9}
}

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    Works referencing / citing this record:

    Nebulin and titin modulate cross-bridge cycling and length-dependent calcium sensitivity
    journal, April 2019

    • Mijailovich, Srboljub M.; Stojanovic, Boban; Nedic, Djordje
    • The Journal of General Physiology, Vol. 151, Issue 5
    • DOI: 10.1085/jgp.201812165