The large amount of data published in the last 10-15 years indicate that myosin isoforms are the major determinant of the large functional heterogeneity of the key contractile and biochemical properties of skeletal muscle fibres, including velocity of shortening, ATP consumption and power. Recent evidences are difficult to reconcile with such an idea and suggest that the properties of muscle fibres that are likely to depend on myosin, such as velocity of shortening, can change without a change in myosin isoform. That a given myosin isoform can modify its properties without shifting to another isoform is confirmed by some analyses of isolated myosin in vitro. The present review is mainly focused on findings that challenge the role of myosin isoforms in determining the functional heterogeneity of skeletal muscle. The work also reports on potential mechanisms behind such changes in myosin function independent of a shift in myosin isoform: the coexistence of different myosin heavy chain (MHC) isoforms in the same fibre, the existence of as yet undetected MHC isoforms, myosin light chain isoforms, post-translational modifications of myosin, the role of other myofibrillar proteins, geometry of the sarcomere and the myosin concentration in single fibres.