Myopathies encompass a wide variety of acquired and hereditary disorders. The pathomechanisms include structural and functional changes affecting, e.g., myofiber metabolism and contractile properties. In this study, we observed increased passive tension (PT) of skinned myofibers from patients with
myofibrillar myopathy (MFM) caused by FLNC mutations (MFM-filaminopathy) and
limb-girdle muscular dystrophy type-2A due to CAPN3 mutations (
LGMD2A), compared to healthy control myofibers. Because the giant
protein titin determines myofiber PT, we measured its molecular size and the
titin-to-
myosin ratio, but found no differences between
myopathies and controls. All-
titin phosphorylation and site-specific phosphorylation in the PEVK region were reduced in
myopathy, which would be predicted to lower PT. Electron microscopy revealed extensive ultrastructural changes in myofibers of various hereditary
myopathies and also suggested massive binding of
proteins to the sarcomeric I-band region, presumably
heat shock proteins (HSPs), which can translocate to elastic
titin under stress conditions. Correlative immunofluorescence and immunoelectron microscopy showed that two small HSPs (HSP27 and αB-
crystallin) and the
ATP-dependent chaperone HSP90 translocated to the
titin springs in
myopathy. The small HSPs, but not HSP90, were upregulated in myopathic versus control muscles. The
titin-binding pattern of chaperones was regularly observed in
Duchenne muscular dystrophy (DMD),
LGMD2A, MFM-filaminopathy, MFM-
myotilinopathy, titinopathy, and inclusion body
myopathy due to mutations in
valosin-containing protein, but not in acquired
sporadic inclusion body myositis. The three HSPs also associated with elastic
titin in mouse models of DMD and MFM-filaminopathy. Mechanical measurements on skinned human myofibers incubated with exogenous small HSPs suggested that the elevated PT seen in
myopathy is caused, in part, by chaperone-binding to the
titin springs. Whereas this interaction may be protective in that it prevents sarcomeric
protein aggregation, it also has detrimental effects on sarcomere function. Thus, we identified a novel pathological phenomenon common to many hereditary
muscle disorders, which involves sarcomeric alterations.