Mechanical parameters of the molecular motor myosin II determined in permeabilised fibres from slow and fast skeletal muscles of the rabbit

• Published in: The Journal of physiology 2018-04, Vol.596 (7), p.1243-1257
• Main Authors: Percario, Valentina, Boncompagni, Simona, Protasi, Feliciano, Pertici, Irene, Pinzauti, Francesca, Caremani, Marco
• Format: Article
• Language: English
• Subjects: Actin; Calcium; Dextran; fast myosin isoform; Isoforms; Muscle; Muscle contraction; muscle mechanics; Musculoskeletal system; Myosin; myosin motor force; myosin motor stiffness; Research Paper; Skeletal muscle; slow myosin isoform
• ISSN: 0022-3751; 1469-7793
• DOI: 10.1113/JP275404

Key points The different performance of slow and fast muscles is mainly attributed to diversity of the myosin heavy chain (MHC) isoform expressed within them. In this study fast sarcomere‐level mechanics has been applied to Ca2+‐activated single permeabilised fibres isolated from soleus (containing the slow myosin isoform) and psoas (containing the fast myosin isoform) muscles of rabbit for a comparative definition of the mechano‐kinetics of force generation by slow and fast myosin isoforms in situ. The stiffness and the force of the slow myosin isoform are three times smaller than those of the fast isoform, suggesting that the stiffness of the myosin motor is a determinant of the isoform‐dependent functional diversity between skeletal muscles. These results open the question of the mechanism that can reconcile the reduced performance of the slow MHC with the higher efficiency of the slow muscle. The skeletal muscle exhibits large functional differences depending on the myosin heavy chain (MHC) isoform expressed in its molecular motor, myosin II. The differences in the mechanical features of force generation by myosin isoforms were investigated in situ by using fast sarcomere‐level mechanical methods in permeabilised fibres (sarcomere length 2.4 μm, temperature 12°C, 4% dextran T‐500) from slow (soleus, containing the MHC‐1 isoform) and fast (psoas, containing the MHC‐2X isoform) skeletal muscle of the rabbit. The stiffness of the half‐sarcomere was determined at the plateau of Ca2+‐activated isometric contractions and in rigor and analysed with a model that accounted for the filament compliance to estimate the stiffness of the myosin motor (ε). ε was 0.56 ± 0.04 and 1.70 ± 0.37 pN nm−1 for the slow and fast isoform, respectively, while the average strain per attached motor (s0) was similar (∼3.3 nm) in both isoforms. Consequently the force per motor (F0 = εs0) was three times smaller in the slow isoform than in the fast isoform (1.89 ± 0.43 versus 5.35 ± 1.51 pN). The fraction of actin‐attached motors responsible for maximum isometric force at saturating Ca2+ (T0,4.5) was 0.47 ± 0.09 in soleus fibres, 70% larger than that in psoas fibres (0.29 ± 0.08), so that F0 in slow fibres was decreased by only 53%. The lower stiffness and force of the slow myosin isoform open the question of the molecular basis of the higher efficiency of slow muscle with respect to fast muscle. Key points The different performance of slow and fast muscles is mainly attributed to di