Bundling of microtubules in vitro by a high molecular weight protein prepared from the squid axon.
A high molecular weight protein has been partially purified from sheaths of squid giant axons. This protein fraction was capable of restoring the membrane excitability of the squid axon which had been destroyed by internal perfusion of microtubule poison, when perfused along with microtubule proteins (Matsumoto et al. (1979) J. Biochem. 86, 1155-1158). This protein, designated as 260 K protein, was purified by gel filtration and Con A-Sepharose affinity chromatography. The apparent molecular weight of the axonal protein was estimated to be 260,000 by electrophoresis in the presence of sodium dodecylsulfate. This protein was revealed to be a glycoprotein. When phosphocellulose-purified tubulin was incubated with 260 K protein at 36 degrees C in the presence of dimethylsulfoxide, turbidity of the solution was much increased. 260 K protein co-sedimented with microtubles assembled from purified tubulin. Light microscopic and electron microscopic observations revealed that the high turbidity was due to bundling of microtubules which was caused by 260 K protein. On the other hand, the effect of this protein on the turbidity increase was not so prominent when microtubules were assembled from microtubule proteins consisting of tubulin and microtubule-associated proteins. High shear and low shear viscometry and co-sedimentation experiments revealed that 260 K protein had little effect on actin polymerization under the same medium conditions as used in tubulin polymerization.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001369:Axons / Q000502:physiology / Q000648:ultrastructure*; D049832:Decapodiformes; D006023:Glycoproteins / Q000302:isolation & purification / Q000502:physiology; D007700:Kinetics; D008854:Microscopy, Electron; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure*; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000502:physiology*
D000596:Amino Acids; D006023:Glycoproteins; D009419:Nerve Tissue Proteins