Actin-caldesmon-myosin-subfragment-1 ternary complex viewed by electron microscopy. Competitive actin binding region for caldesmon and myosin subfragment-1.
An earlier electron microscopic study using different caldesmon forms complexed with actin revealed that the aggregates produced display regular periodic striation after antibody labeling of the 35-kDa caldesmon fragment. This approach provides further evidence that a caldesmon fragment, even as small as 15 kDa, can induce actin filaments to assemble into bundles. The observed difference in the compactness of these structures, depending on the use of the 15-kDa fragment instead of the 35-kDa fragment, suggests the existence of more than one actin-binding site in the caldesmon molecule. In this study, the caldesmon-induced process of F-actin association was investigated in the presence of skeletal myosin subfragment-1, using light-scattering methods, cosedimentation experiment and electron microscopic techniques. We show that the actin-caldesmon association is partially destabilized in the presence of subfragment-1 and this leads to a ternary complex formation. Immunogold labelling of the actin filaments still reveals the presence of caldesmon within this structure. This latter result strengthens the hypothesis that actin has a site(s) able to bind both caldesmon and myosin subfragment-1, as detected by recent NMR observations. This evidence is discussed with respect to the regulatory function of caldesmon during smooth muscle contraction.
Harricane
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MC
Centre de Recherche de Biochimie Macromoléculaire du CNRS, INSERM U.249, Université de Montpellier, France.
Bonet-Kerrache
A
A
Cavadore
C
C
Mornet
D
D
D000199:Actins / Q000737:chemistry* / Q000648:ultrastructure; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002148:Calmodulin-Binding Proteins / Q000737:chemistry* / Q000648:ultrastructure; D008854:Microscopy, Electron; D015879:Myosin Subfragments / Q000737:chemistry* / Q000648:ultrastructure; D011487:Protein Conformation
D000199:Actins; D002148:Calmodulin-Binding Proteins; D015879:Myosin Subfragments