Extensive purification from Acanthamoeba castellanii of a microtubule-dependent translocator with microtubule-activated Mg2+-ATPase activity.

B. Kachar, J. P. Albanesi, H. Fujisaki, E. D. Korn

Research output: Contribution to journalArticle

15 Scopus citations

Abstract

A protein which supported MgATP-dependent movement of latex beads from the minus to the plus end of microtubules and which had microtubule-activated Mg2+-ATPase was purified from Acanthamoeba castellanii. At concentrations as low as 0.6 micrograms ml-1, the translocator supported movement of beads at a rate of 3 to 4 micron s-1. The translocator protein had a Ca2+-ATPase activity of 1.7 mumol min-1 mg-1 and a Mg2+-ATPase activity of about 0.03 mumol min-1 mg-1 in the absence of microtubules. The Mg2+-ATPase in the presence of microtubules had a Vmax of 3.4 mumol min-1 mg-1; half-maximal Mg2+-ATPase activity required only 0.45 microM microtubules (concentration of dimer subunits). The highly purified native protein had a Stokes radius of 8.5 nm, and three polypeptides of Mr 134,000, 139,000, and 147,000 were associated with the fractions that had maximum translocator and ATPase activities.

Original languageEnglish (US)
Pages (from-to)16180-16185
Number of pages6
JournalThe Journal of biological chemistry
Volume262
Issue number33
StatePublished - Nov 25 1987

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Extensive purification from Acanthamoeba castellanii of a microtubule-dependent translocator with microtubule-activated Mg2+-ATPase activity.'. Together they form a unique fingerprint.

Cite this