Interactions of enolase isoforms with tubulin and microtubules during myogenesis

• Published in: Biochimica et biophysica acta Vol. 1770; no. 6; pp. 919 - 926
• Main Authors: Keller, A., Peltzer, J., Carpentier, G., Horváth, I., Oláh, J., Duchesnay, A., Orosz, F., Ovádi, J.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.06.2007; Elsevier
• Subjects: Animals; Cell Differentiation; Cell Differentiation - physiology; Cells, Cultured; Cellular Biology; Confocal microscopy; Enolase isoform; Enzyme-Linked Immunosorbent Assay; Glyceric Acids; Glyceric Acids - chemistry; Glyceric Acids - metabolism; Glycolysis; Isoenzymes; Isoenzymes - chemistry; Isoenzymes - metabolism; Life Sciences; Mice; Microtubule; Microtubules; Microtubules - chemistry; Microtubules - enzymology; Muscle Development; Muscle Development - physiology; Muscle Fibers; Muscle Fibers, Skeletal - cytology; Muscle Fibers, Skeletal - enzymology; Muscle, Skeletal; Muscle, Skeletal - cytology; Muscle, Skeletal - metabolism; Myogenesis; Phosphopyruvate Hydratase; Phosphopyruvate Hydratase - chemistry; Phosphopyruvate Hydratase - metabolism; Protein Binding; Protein Binding - physiology; Protein Transport; Protein Transport - physiology; Satellite Cells, Skeletal Muscle; Satellite Cells, Skeletal Muscle - cytology; Satellite Cells, Skeletal Muscle - enzymology; Surface Plasmon Resonance; Tubulin; Tubulin - chemistry; Tubulin - metabolism; Glycolysis; Confocal microscopy; Enolase isoform; Tubulin; Microtubule; Myogenesis
• ISSN: 0304-4165; 0006-3002; 0167-4889; 1872-8006
• DOI: 10.1016/j.bbagen.2007.01.015

Enolase is a glycolytic enzyme, expressed as cell-type specific isoforms in higher vertebrates. Herein we demonstrated for the first time that enolase isoforms interact with microtubules during muscle satellite cell differentiation. While in undifferentiated myoblasts the ubiquitous αα enolase isoform, expressed at high level, exhibited extensive co-localization with microtubules, the muscle-specific ββ isoform, expressed at low level, did not. During differentiation, the level of β subunit increased significantly; the α and β enolase immunoreactivities were detected both in cytosol and along the microtubules. We identified tubulin from muscle extract as an interacting protein for immobilized ββ enolase. ELISA and surface plasmon resonance measurements demonstrated the direct binding of enolase isoforms to tubulin with an apparent K D below the micromolar range, and indicated that the presence of 0.8 mM 2-phosphoglycerate abolished the interaction. Our data showed that, at various stages of myogenic differentiation, microtubules were decorated by different enolase isoforms, which was controled by the abundance of both partners. We suggest that the binding of enolase to microtubules could contribute to the regulation of the dynamism of the cytoskeletal filaments known to occur during the transition from myoblast to myotubes.