Structure and dynamics of the kinesin-microtubule interaction revealed by fluorescence polarization microscopy

Hernando Sosa, Ana B. Asenjo, Erwin J.G. Peterman

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Fluorescence polarization microscopy (FPM) is the analysis of the polarization of light in a fluorescent microscope in order to determine the angular orientation and rotational mobility of fluorescent molecules. Key advantages of FPM, relative to other structural analysis techniques, are that it allows the detection of conformational changes of fluorescently labeled macromolecules in real time in physiological conditions and at the single-molecule level. In this chapter we describe in detail the FPM experimental set-up and analysis methods we have used to investigate structural intermediates of the motor protein kinesin-1 associated with its walking mechanism along microtubules. We also briefly describe additional FPM methods that have been used to investigate other macromolecular complexes.

Original languageEnglish (US)
Pages (from-to)505-519
Number of pages15
JournalMethods in Cell Biology
Volume95
Issue numberC
DOIs
StatePublished - Jan 1 2010

ASJC Scopus subject areas

  • Cell Biology

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