Pseudo-phosphorylation of essential light chains affects the functioning of skeletal muscle myosin

• Published in: Biophysical chemistry 2023-01, Vol.292, p.106936-106936, Article 106936
• Main Authors: Yampolskaya, Daria S., Kopylova, Galina V., Shchepkin, Daniil V., Nabiev, Salavat R., Nikitina, Larisa V., Walklate, Jonathan, Ziganshin, Rustam H., Bershitsky, Sergey Y., Geeves, Michael A., Matyushenko, Alexander M., Levitsky, Dmitrii I.
• Format: Article
• Language: English
• Subjects: Actin-myosin interaction; Actins; Essential light chain; Mechanism of muscle contraction; Muscle, Skeletal; Myosin; Myosins; Phosphorylation; Pseudo-phosphorylation; Skeletal Muscle Myosins
• ISSN: 0301-4622; 1873-4200
• DOI: 10.1016/j.bpc.2022.106936

The work aimed to investigate how the phosphorylation of the myosin essential light chain of fast skeletal myosin (LC1) affects the functional properties of the myosin molecule. Using mass-spectrometry, we revealed phosphorylated peptides of LC1 in myosin from different fast skeletal muscles. Mutations S193D and T65D that mimic natural phosphorylation of LC1 were produced, and their effects on functional properties of the entire myosin molecule and isolated myosin head (S1) were studied. We have shown that T65D mutation drastically decreased the sliding velocity of thin filaments in an in vitro motility assay and strongly increased the duration of actin-myosin interaction in optical trap experiments. These effects of T65D mutation in LC1 observed only with the whole myosin but not with S1 were prevented by double T65D/S193D mutation. The T65D and T65D/S193D mutations increased actin-activated ATPase activity of S1 and decreased ADP affinity for the actin-S1 complex. The results indicate that pseudo-phosphorylation of LC1 differently affects the properties of the whole myosin molecule and its isolated head. Also, the results show that phosphorylation of LC1 of skeletal myosin could be one more mechanism of regulation of actin-myosin interaction that needs further investigation. [Display omitted] •Essential light chain (LC1) of skeletal myosin can be phosphorylated.•Mutations S193D and T65D mimic phosphorylation of LC1•T65D mutation differently affects properties of the whole myosin molecule and its isolated head (S1).•Double T65D/S193D mutation prevents dramatic effects of T65D mutation on myosin properties.•LC1 phosphorylation could be an extra mechanism of regulation of actin-myosin interaction.