Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Methods Enzymol
2014 Jan 01;540:339-60. doi: 10.1016/B978-0-12-397924-7.00019-4.
Show Gene links
Show Anatomy links
Micropattern-guided assembly of overlapping pairs of dynamic microtubules.
Fourniol FJ
,
Li TD
,
Bieling P
,
Mullins RD
,
Fletcher DA
,
Surrey T
.
???displayArticle.abstract???
Interactions between antiparallel microtubules are essential for the organization of spindles in dividing cells. The ability to form immobilized antiparallel microtubule pairs in vitro, combined with the ability to image them via TIRF microscopy, permits detailed biochemical characterization of microtubule cross-linking proteins and their effects on microtubule dynamics. Here, we describe methods for chemical micropatterning of microtubule seeds on glass surfaces in configurations that specifically promote the formation of antiparallel microtubule overlaps in vitro. We demonstrate that this assay is especially well suited for reconstitution of minimal midzone overlaps stabilized by the antiparallel microtubule cross-linking protein PRC1 and its binding partners. The micropatterning method is suitable for use with a broad range of proteins, and the assay is generally applicable to any microtubule cross-linking protein.
Aoyama,
Self-organized optical device driven by motor proteins.
2013, Pubmed
Aoyama,
Self-organized optical device driven by motor proteins.
2013,
Pubmed
Bieling,
A minimal midzone protein module controls formation and length of antiparallel microtubule overlaps.
2010,
Pubmed
,
Xenbase
Braun,
Adaptive braking by Ase1 prevents overlapping microtubules from sliding completely apart.
2011,
Pubmed
Castoldi,
Purification of brain tubulin through two cycles of polymerization-depolymerization in a high-molarity buffer.
2003,
Pubmed
Duellberg,
End-binding proteins and Ase1/PRC1 define local functionality of structurally distinct parts of the microtubule cytoskeleton.
2013,
Pubmed
Gaillard,
Two microtubule-associated proteins of Arabidopsis MAP65s promote antiparallel microtubule bundling.
2008,
Pubmed
Gell,
Microtubule dynamics reconstituted in vitro and imaged by single-molecule fluorescence microscopy.
2010,
Pubmed
Ghosh,
Micropattern-controlled local microtubule nucleation, transport, and mesoscale organization.
2013,
Pubmed
Glotzer,
The 3Ms of central spindle assembly: microtubules, motors and MAPs.
2009,
Pubmed
Hentrich,
Microtubule organization by the antagonistic mitotic motors kinesin-5 and kinesin-14.
2010,
Pubmed
,
Xenbase
Hyman,
Preparation of modified tubulins.
1991,
Pubmed
Janson,
Crosslinkers and motors organize dynamic microtubules to form stable bipolar arrays in fission yeast.
2007,
Pubmed
Kapitein,
The bipolar mitotic kinesin Eg5 moves on both microtubules that it crosslinks.
2005,
Pubmed
,
Xenbase
Mollinari,
PRC1 is a microtubule binding and bundling protein essential to maintain the mitotic spindle midzone.
2002,
Pubmed
Nunes Bastos,
Aurora B suppresses microtubule dynamics and limits central spindle size by locally activating KIF4A.
2013,
Pubmed
Portran,
Quantification of MAP and molecular motor activities on geometrically controlled microtubule networks.
2013,
Pubmed
Roostalu,
Directional switching of the kinesin Cin8 through motor coupling.
2011,
Pubmed
Shaner,
Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein.
2004,
Pubmed
Su,
Microtubule-sliding activity of a kinesin-8 promotes spindle assembly and spindle-length control.
2013,
Pubmed
Subramanian,
Insights into antiparallel microtubule crosslinking by PRC1, a conserved nonmotor microtubule binding protein.
2010,
Pubmed
Subramanian,
Marking and measuring single microtubules by PRC1 and kinesin-4.
2013,
Pubmed
Sørensen,
A favorable solubility partner for the recombinant expression of streptavidin.
2003,
Pubmed
Telley,
Reconstitution and quantification of dynamic microtubule end tracking in vitro using TIRF microscopy.
2011,
Pubmed
Vernì,
Feo, the Drosophila homolog of PRC1, is required for central-spindle formation and cytokinesis.
2004,
Pubmed
Waichman,
Maleimide photolithography for single-molecule protein-protein interaction analysis in micropatterns.
2011,
Pubmed
van den Wildenberg,
The homotetrameric kinesin-5 KLP61F preferentially crosslinks microtubules into antiparallel orientations.
2008,
Pubmed
