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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: Publikationen MPI-CBG 2010-arch     Display Documents



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ID: 546679.0, MPI für molekulare Zellbiologie und Genetik / Publikationen MPI-CBG 2010-arch
Studying kinesin motors by optical 3D-nanometry in gliding motility assays
Authors:Nitzsche, Bert; Bormuth, Volker; Bräuer, Corina; Howard, Jonathon; Ionov, Leonid; Kerssemakers, Jacob W. J.; Korten, Till; Leduc, Cecile; Ruhnow, Felix; Diez, Stefan
Place of Publication:Amsterdam, Netherlands
Publisher:Elsevier
Date of Publication (YYYY-MM-DD):2010
Title of Book:Microtubules, in vitro
Start Page:247
End Page:271
Sequence Number of Inbook:14
Full Name of Book-Editor(s):Wilson, Leslie
Copyright:not available
Review Status:not specified
Audience:Experts Only
Intended Educational Use:No
Abstract / Description:Recent developments in optical microscopy and nanometer tracking have facilitated our understanding of microtubules and their associated proteins. Using fluorescence microscopy, dynamic interactions are now routinely observed in vitro on the level of single molecules, mainly using a geometry in which labeled motors move on surface-immobilized microtubules. Yet, we think that the historically older gliding geometry, in which motor proteins bound to a substrate surface drive the motion microtubules, offers some unique advantages. (1) Motility can be precisely followed by coupling multiple fluorophores and/or single bright labels to the surface of microtubules without disturbing the activity of the motor proteins. (2) The number of motor proteins involved in active transport can be determined by several strategies. (3) Multimotor studies can be performed over a wide range of motor densities. These advantages allow for studying cooperativity of processive as well as nonprocessive motors. Moreover, the gliding geometry has proven to be most promising for nanotechnological applications of motor proteins operating in synthetic environments. In this chapter we review recent methods related to gliding motility assays in conjunction with 3D-nanometry. In particular, we aim to provide practical advice on how to set up gliding assays, how to acquire high-precision data from microtubules and attached quantum dots, and how to analyze data by 3D-nanometer tracking.
External Publication Status:published
Document Type:InBook
Communicated by:nn
Affiliations:MPI für molekulare Zellbiologie und Genetik
Identifiers:LOCALID:4131
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