Interactions of enolase isoforms with tubulin and microtubules during myogenesis.
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 alphaalpha enolase isoform, expressed at high level, exhibited extensive co-localization with microtubules, the muscle-specific betabeta isoform, expressed at low level, did not. During differentiation, the level of beta subunit increased significantly; the alpha and beta enolase immunoreactivities were detected both in cytosol and along the microtubules. We identified tubulin from muscle extract as an interacting protein for immobilized betabeta enolase. ELISA and surface plasmon resonance measurements demonstrated the direct binding of enolase isoforms to tubulin with an apparent KD 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 controlled 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.
Keller
A
A
Laboratoire CRRET, CNRS UMR 7149, Université Paris 12, 94010 Créteil, France. keller@univ-paris12.fr
Peltzer
J
J
Carpentier
G
G
Horváth
I
I
Oláh
J
J
Duchesnay
A
A
Orosz
F
F
Ovádi
J
J
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005988:Glyceric Acids / Q000737:chemistry / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008870:Microtubules / Q000737:chemistry / Q000201:enzymology*; D024510:Muscle Development / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000201:enzymology; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology / Q000201:enzymology*; D020349:Surface Plasmon Resonance; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
D005988:Glyceric Acids; D007527:Isoenzymes; D014404:Tubulin; C008885:2-phosphoglycerate; D010751:Phosphopyruvate Hydratase