Abstract
Kinesin was purified from bovine adrenal medulla. The sedimentation coefficient was 8.8 S. Sedimentation equilibrium ultracentrifugation studies showed the molecular weight of kinesin to be 300,000. The calculated axial ratio was 1:16. The Stokes radius was estimated to be 8.9 nm by gel filtration. Circular dichroism showed the alpha-helix content to be about 50%. Purified kinesin preparation contained a major polypeptide with a molecular weight of 120,000 and minor ones with molecular weights of 71,000, 68,000, and 65,000. Bovine adrenal kinesin had an ATPase activity which was stimulated severalfold by microtubules to a specific activity of about 0.1 mumol/min.mg. Kinesin molecules adsorbed to a glass slide promoted the movement of microtubules on the glass surface at a rate of about 0.5 micron/s. Immunostaining of EBTr (bovine embryonic trachea fibroblast) cells and bovine adrenal chromaffin cells in interphase with an affinity-purified antibody against the major polypeptide of kinesin showed that some kinesin was located on microtubules and the rest distributed throughout the cytoplasm in a diffuse manner. EBTr cells in mitotic phase gave a staining pattern showing that kinesin was present throughout the cytoplasm with higher concentration in the region of mitotic apparatus.
Highlights
From the Departmentof Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Bunkyo-ku, Tokyo113, Japan and the §LifeScience Institute, Sophia Uniuersity, Kioi-cho, Toky1o02,Japan
Kinesin was purified from bovine adrenal medulla. microtubules in the presence of AMP-PNP’ and ATP, re
We determined the physical properties of kinesin including molecular weight, sedimentation coefficient, Stokes radius, and a-helix content.We determined the catalytic nature of adrenal kinesin, such as ATPase activity and an ability to promote the movement of microtubules on the surface of a slide glass
Summary
Hiromu Murofushi, AtsushiIkai, Koji Okuhara, Susumu KotaniS, HiroyukAi izawa, Konosuke Kumakurat, and Hikoichi Sakai. Immunostaining of EBTr (bovine embryonic tracheaof Kuznetsov and Gelfand [14]bovine brain kinesin has a fibroblast)cells and bovine adrenal chromaffcinells in microtubule-stimulated Mg+-ATPasewith a specific activity interphase with an affinity-purified antibody against higher than that of dynein. A protein termed kinesin, which mediates microtubulebased transport of latex beads, was purified from axoplasm of squid giant axons, squid optic lobes, and mammalian brains [7], taking advantage of the higher and lower affinity to cholamines by chromaffin cells has been used as the model system for the study on exocytosis of neurotransmitters by neurons. It is important to study kinesin in adrenal medulla in detail
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
