Microtubule assembly of isotypically purified tubulin and its mixtures.

Abstract:

:Numerous isotypes of the structural protein tubulin have now been characterized in various organisms and their expression offers a plausible explanation for observed differences affecting microtubule function in vivo. While this is an attractive hypothesis, there are only a handful of studies demonstrating a direct influence of tubulin isotype composition on the dynamic properties of microtubules. Here, we present the results of experimental assays on the assembly of microtubules from bovine brain tubulin using purified isotypes at various controlled relative concentrations. A novel data analysis is developed using recursive maps which are shown to be related to the master equation formalism. We have found striking similarities between the three isotypes of bovine tubulin studied in regard to their dynamic instability properties, except for subtle differences in their catastrophe frequencies. When mixtures of tubulin isotypes are analyzed, their nonlinear concentration dependence is modeled and interpreted in terms of lower affinities of tubulin dimers belonging to the same isotype than those that represent different isotypes indicating hitherto unsuspected influences of tubulin dimers on each other within a microtubule. Finally, we investigate the fluctuations in microtubule assembly and disassembly rates and conclude that the inherent rate variability may signify differences in the guanosine-5'-triphosphate composition of the growing and shortening microtubule tips. It is the main objective of this article to develop a quantitative model of tubulin polymerization for individual isotypes and their mixtures. The possible biological significance of the observed differences is addressed.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Rezania V,Azarenko O,Jordan MA,Bolterauer H,Ludueña RF,Huzil JT,Tuszynski JA

doi

10.1529/biophysj.108.132233

subject

Has Abstract

pub_date

2008-08-01 00:00:00

pages

1993-2008

issue

4

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(08)70157-7

journal_volume

95

pub_type

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