A porcine brain protein (35 K protein) which bundles microtubules and its identification as glyceraldehyde 3-phosphate dehydrogenase.
A protein which binds to both tubulin and tubulin polymer was isolated from porcine brains. This protein has a molecular weight of 35,000 on SDS-polyacrylamide gel electrophoresis (designated as 35 K protein). The 35 K protein was purified through several steps of purification including ammonium sulfate fractionation, Sephadex G-100 gel filtration column chromatography, microtubule protein-agarose gel affinity column chromatography and phosphocellulose column chromatography. The 35 K protein caused pronounced enhancement of the turbidity increase produced by tubulin polymerization in the presence of DMSO, but did not have the ability to initiate polymerization of pure tubulin in the absence of DMSO. It was demonstrated that 35 K protein co-sediments with tubulin polymer in a concentration-dependent manner. Electron microscopic observation revealed the formation of bundles of tubulin polymer. Since the effect of 35 K protein was coupled with tubulin polymerization, 35 K protein did not cause the turbidity increase under conditions where tubulin polymerization was inhibited by Ca2+ or colchicine. The 35 K protein adsorbed on tubulin-Sepharose 4B was eluted by the addition of 2 mM ATP. ATP was shown to inhibit the interaction of 35 K protein with tubulin dimer or polymer. The 35 K protein was finally identified as glyceraldehyde 3-phosphate dehydrogenase from properties such as mobility on SDS-polyacrylamide gel electrophoresis, cleavage pattern on limited proteolysis, ability to bind to tubulin, and so on.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D001923:Brain Chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008970:Molecular Weight; D011108:Polymers / Q000378:metabolism; D013552:Swine; D014404:Tubulin / Q000378:metabolism
D011108:Polymers; D014404:Tubulin; D000255:Adenosine Triphosphate; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases