Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

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

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [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)
+ Show Author Affiliations
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.
Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
NIH; National Inst. of General Medical Sciences (NIGMS); USDOE
Grant/Contract Number:
AC02-06CH11357
OSTI ID:
1478092
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 41 Vol. 115; ISSN 0027-8424
Publisher:
National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
ENGLISH

References (44)

Nebulin is a full-length template of actin filaments in the skeletal muscle sarcomere: an immunoelectron microscopic study of its orientation and span with site-specific monoclonal antibodies journal October 1993
Nebulin as a giant actin-binding template protein in skeletal muscle sarcomere Interaction of actin and cloned human nebulin fragments journal April 1991
Evidence that nebulin is a protein-ruler in muscle thin filaments journal May 1991
Myosin Heads Contribute to the Maintenance of Filament Order in Relaxed Rabbit Muscle journal September 2010
Thick-Filament Strain and Interfilament Spacing in Passive Muscle: Effect of Titin-Based Passive Tension journal March 2011
Altered myofilament function depresses force generation in patients with nebulin-based nemaline myopathy (NEM2) journal May 2010
Nemaline myopathy caused by mutations in the nebulin gene may present as a distal myopathy journal August 2011
The Nebulin family: an actin support group journal January 2011
Equatorial x-ray diffraction from single skinned rabbit psoas fibers at various degrees of activation. Changes in intensities and lattice spacing journal November 1985
Time-resolved x-ray diffraction studies on the intensity changes of the 5.9 and 5.1 nm actin layer lines from frog skeletal muscle during an isometric tetanus using synchrotron radiation journal June 1985
Time-resolved x-ray diffraction study of the troponin-associated reflexions from the frog muscle journal September 1992
X-ray diffraction evidence for the extensibility of actin and myosin filaments during muscle contraction journal December 1994
Analysis of equatorial x-ray diffraction patterns from muscle fibers: factors that affect the intensities journal May 1995
The complete primary structure of human nebulinand its correlation to muscle structure journal January 1995
X-ray Diffraction from Living Striated Muscle during Contraction journal June 1965
Nebulin regulates thin filament length, contractility, and Z-disk structure in vivo journal August 2006
A myopathy-linked tropomyosin mutation severely alters thin filament conformational changes during activation journal May 2010
Architecture of the sarcomere matrix of skeletal muscle: immunoelectron microscopic evidence that suggests a set of parallel inextensible nebulin filaments anchored at the Z line journal December 1988
Three-dimensional stochastic model of actin–myosin binding in the sarcomere lattice journal November 2016
Deleting exon 55 from the nebulin gene induces severe muscle weakness in a mouse model for nemaline myopathy journal May 2013
Thin filament length dysregulation contributes to muscle weakness in nemaline myopathy patients with nebulin deficiency journal April 2009
Nebulin deficiency in adult muscle causes sarcomere defects and muscle-type-dependent changes in trophicity: novel insights in nemaline myopathy journal June 2015
Disrupted myosin cross-bridge cycling kinetics triggers muscle weakness in nebulin-related myopathy journal June 2011
The Filament Lattice of Striated Muscle journal April 1998
Nebulin is a full-length template of actin filaments in the skeletal muscle sarcomere: an immunoelectron microscopic study of its orientation and span with site-specific monoclonal antibodies journal October 1993
Nebulin as a giant actin-binding template protein in skeletal muscle sarcomere Interaction of actin and cloned human nebulin fragments journal April 1991
Evidence that nebulin is a protein-ruler in muscle thin filaments journal May 1991
Understanding the functions of titin and nebulin journal July 1992
Equatorial x-ray diffraction from single skinned rabbit psoas fibers at various degrees of activation. Changes in intensities and lattice spacing journal November 1985
Time-resolved x-ray diffraction studies on the intensity changes of the 5.9 and 5.1 nm actin layer lines from frog skeletal muscle during an isometric tetanus using synchrotron radiation journal June 1985
Time-resolved x-ray diffraction study of the troponin-associated reflexions from the frog muscle journal September 1992
X-ray diffraction measurements of the extensibility of actin and myosin filaments in contracting muscle journal December 1994
X-ray diffraction evidence for the extensibility of actin and myosin filaments during muscle contraction journal December 1994
Analysis of equatorial x-ray diffraction patterns from muscle fibers: factors that affect the intensities journal May 1995
Temperature-induced structural changes in the myosin thick filament of skinned rabbit psoas muscle journal November 1997
The complete primary structure of human nebulinand its correlation to muscle structure journal January 1995
Each Actin Subunit Has Three Nebulin Binding Sites journal March 2002
Myopathy-inducing mutation H40Y in ACTA1 hampers actin filament structure and function journal August 2016
Force generation by skeletal muscle is controlled by mechanosensing in myosin filaments journal November 2015
Direct measurement of stiffness of single actin filaments with and without tropomyosin by in vitro nanomanipulation. journal December 1994
Flexibility of actin filaments derived from thermal fluctuations. Effect of bound nucleotide, phalloidin, and muscle regulatory proteins journal May 1995
Nebulin Alters Cross-bridge Cycling Kinetics and Increases Thin Filament Activation: A NOVEL MECHANISM FOR INCREASING TENSION AND REDUCING TENSION COST journal September 2009
Nebulin as a length regulator of thin filaments of vertebrate skeletal muscles: correlation of thin filament length, nebulin size, and epitope profile journal October 1991
The contributions of filaments and cross-bridges to sarcomere compliance in skeletal muscle: Myofilament and cross-bridge compliance journal September 2014

Cited By (9)

Nebulin and titin modulate cross-bridge cycling and length-dependent calcium sensitivity journal April 2019
Elastic domains of giant proteins in striated muscle: Modeling compliance with rulers journal April 2019
Omecamtiv mecarbil lowers the contractile deficit in a mouse model of nebulin-based nemaline myopathy journal November 2019
Functional Characterization of the Intact Diaphragm in a Nebulin-Based Nemaline Myopathy (NM) Model-Effects of the Fast Skeletal Muscle Troponin Activator tirasemtiv journal October 2019
Estimation of Forces on Actin Filaments in Living Muscle from X-ray Diffraction Patterns and Mechanical Data journal November 2019
Nebulin: big protein with big responsibilities journal January 2020
Nebulin no longer nebulous journal October 2018
KBTBD13 is an actin-binding protein that modulates muscle kinetics journal December 2019
Nebulin nemaline myopathy recapitulated in a compound heterozygous mouse model with both a missense and a nonsense mutation in Neb journal February 2020

Similar Records

Triggering typical nemaline myopathy with compound heterozygous nebulin mutations reveals myofilament structural changes as pathomechanism
Journal Article · Sun May 31 20:00:00 EDT 2020 · Nature Communications · OSTI ID:1631850
Comparative decline of the protein profiles of nebulin in response to denervation in skeletal muscle
Journal Article · Fri Oct 09 00:00:00 EDT 2015 · Biochemical and Biophysical Research Communications · OSTI ID:22592748
Nebulin and titin modulate cross-bridge cycling and length-dependent calcium sensitivity
Journal Article · Wed Apr 03 20:00:00 EDT 2019 · Journal of General Physiology · OSTI ID:1558308

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
X-ray diffraction
muscle biology
physiology
skeletal myopathy