title: Reduction in microtubule dynamics in vitro by brain microtubule-associated proteins and by a microtubule-associated protein-2 second repeated sequence analogue.
abstract: Microtubule-associated protein (MAP) binding to assembled microtubules (MTs) can be reduced by the addition of polyglutamate without significant MT depolymerization or interference with MT elongation reactions. Ensuing polymer length redistribution in MAP-depleted MTs occurs on a time scale characteristic of that observed with MAP-free MTs. The redistribution phase occurs even in the absence of mechanical shearing and without appreciable effects from end-to-end annealing, as indicated by the time course of incremental changes in polymer length and MT number concentration. We also observed higher rates of MT length redistribution when the [MAP]/[tubulin] ratio was decreased. Together, these results demonstrate that MT length redistribution rates are greatly influenced by MAP content, and the data are compatible with the dynamic instability model. We also found that a peptide analogue corresponding to the second repeated sequence in the MT-binding region of MAP-2 can also markedly retard MT length redistribution kinetics, a finding that accords with the ability of this peptide to promote tubulin polymerization in the absence of MAPs and to displace MAP-2 from MTs. These results provide further evidence that MAPs can modulate MT assembly/disassembly dynamics and that peptide analogues can mimic the action of intact MAPs without the need for three contiguous repeated sequences in the MT-binding region.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville.
authors.forename: P S
authors.initials: PS
authors.lastname: Yamauchi
authors.forename: G C
authors.initials: GC
authors.lastname: Flynn
authors.forename: R L
authors.initials: RL
authors.lastname: Marsh
authors.forename: D L
authors.initials: DL
authors.lastname: Purich
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology / Q000502:physiology*; D008870:Microtubules / Q000187:drug effects / Q000502:physiology*; D008969:Molecular Sequence Data; D011099:Polyglutamic Acid / Q000494:pharmacology; D012091:Repetitive Sequences, Nucleic Acid
chemical_list: D008869:Microtubule-Associated Proteins; D011099:Polyglutamic Acid