title: Ion specific effects in bundling and depolymerization of taxol-stabilized microtubules.
abstract: Microtubules (MTs) are nanometer scale hollow cylindrical biological polyelectrolytes. They are assembled from alpha/beta-tubulin dimers, which stack to form protofilaments (PFs) with lateral interactions between PFs resulting in the curved MT. In cells, MTs and their assemblies are critical components in a range of functions from providing tracks for the transport of cargo to forming the spindle structure during mitosis. Previous studies have, shown that while cations with valence equal to or larger than 3+ tend to assemble tight 3D bundles of taxol-stabilized MTs, certain divalent cations induce relatively loose 2D bundles of different symmetry (D. J. Needleman et al., Proc. Natl. Acad. Sci. U. S. A., 2004, 101, 16099). Similarly, divalent cations form 2D bundles of DNA adsorbed on cationic membranes (I. Koltover et al., Proc. Natl. Acad. Sci. U. S. A., 2000, 97, 14046). The bundling behavior for these biological polyelectrolyte systems is qualitatively in agreement with current theory. Here, we present results which show that, unlike the case for DNA adsorbed on cationic membranes, bundling of taxol-stabilized MTs occurs only for certain divalent cations above a critical ion concentration (e.g. Ca2+, Sr2+, Ba2+). Instead, many divalent cations pre-empt the bundling transition and depolymerize taxol-stabilized MTs at a lower counterion concentration. Although previous cryogenic TEM has shown that, in the absence of taxol, Ca2+ depolymerizes MTs assembling in buffers containing GTP (guanosine triphosphate), our finding is surprising given the know stabilizing effects of taxol on GDP (guanosine diphosphate)-MTs. The ion concentration required for MT depolymerization decreases with increasing atomic number for the divalents Mg2+, Mn2+, Co2+, and Zn2+. GdCl3 (3+) is found to be extremely efficient at MT depolymerization requiring ion concentrations of about 1 mM, while oligolysine(2+), is observed not to depolymerize MTs at concentrations as high as 144 mM. The surprising MT depolymerization results are discussed in the context of divalents either disrupting lateral interactions between PFs (which are strengthened for taxol containing beta-tubulin) or interfering with taxol's ability to induce flexibility at the interface between two tubulin dimers in the same PF (which has been recently suggested as a mechanism by which taxol stabilizes MTs post-hydrolysis with the induced flexibility counteracting the kink between GDP-tublin dimers in a PF).
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Needleman
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Ojeda-Lopez
authors.forename: Uri
authors.initials: U
authors.lastname: Raviv
authors.forename: Herbert P
authors.initials: HP
authors.lastname: Miller
authors.forename: Youli
authors.initials: Y
authors.lastname: Li
authors.forename: Chaeyeon
authors.initials: C
authors.lastname: Song
authors.forename: Stuart C
authors.initials: SC
authors.lastname: Feinstein
authors.forename: Leslie
authors.initials: L
authors.lastname: Wilson
authors.forename: Myung Chul
authors.initials: MC
authors.lastname: Choi
authors.forename: Cyrus R
authors.initials: CR
authors.lastname: Safinya
mesh_terms: D001704:Biopolymers / Q000737:chemistry*; D002413:Cations, Divalent; D046529:Microscopy, Electron, Transmission; D008870:Microtubules / Q000737:chemistry / Q000187:drug effects*; D017239:Paclitaxel / Q000494:pharmacology*; D053838:Scattering, Small Angle
chemical_list: D001704:Biopolymers; D002413:Cations, Divalent; D017239:Paclitaxel