Fluorescent Methods for Molecular Motors

306 Pages·2014·7.952 MB·English

by  Christopher P. Toseland, Natalia Fili (eds.)

Experientia Supplementum 105
Christopher P. Toseland
Natalia Fili Editors
Fluorescent
Methods for
Molecular
Motors
Experientia Supplementum
Volume 105
For furthervolumes:
http://www.springer.com/series/4822
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Christopher P. Toseland (cid:129) Natalia Fili
Editors
Fluorescent Methods for
Molecular Motors
Editors
ChristopherP.Toseland NataliaFili
ChromosomeOrganisationandDynamics DepartmentofCellularPhysiology
Max-PlanckInstituteofBiochemistry Ludwig-Maximilians-Universita¨tMu¨nchen
Martinsried Munich
Germany Germany
ISSN1023-294X
ISBN978-3-0348-0855-2 ISBN978-3-0348-0856-9(eBook)
DOI10.1007/978-3-0348-0856-9
SpringerBaselHeidelbergDordrechtNewYorkLondon
LibraryofCongressControlNumber:2014947103
©SpringerBasel2014
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For Thalia...You remind us of how much we
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Preface
The term “Fluorescence” was initially coined by George Gabriel Stokes in 1852,
butthephenomenonhadalreadybeenrecordedforover300years.Ourknowledge
of fluorescence expanded through the pioneering work of Perrin, Jablonski and
Fo¨rster and Weber. This work coincided with the synthesis of organic dyes—the
fluorophores. Our ways to manipulate fluorophores have grown to suit our needs
and our ways to detect them have developed more than could be imagined a few
yearsago.Almostalllaboratoriesutilizefluorescence,whetheritismicroscopyor
asareagent...wecouldconsiderthatwenowliveinthe“AgeofFluorescence”.
Fluorescence and motor proteins have a long interconnected relationship.
Indeed,theprogressinfluorescencemethodshasdirectlyadvancedourknowledge
ofthesemolecularmachines.
Writtenbyexperts,thisbookfocusesontheapplicationoffluorescencetostudy
motor proteins. It is intended for a large community of biochemists, biophysicists
and cell biologists who study a diverse collection of motor proteins. It can be
usedbyresearcherstogainaninsightintotheirfirstexperiments,orbyexperienced
researchers who are looking to expand their research to new areas. Within each
chapter you will find valuable advice for executing the experiments, along with
detailedbackgroundknowledgeinordertodevelopyourownexperiments.
Wethankalltheauthorsfortheirhardworkmakingthisbookareality.
Martinsried,Germany ChristopherP.Toseland
Munich,Germany NataliaFili
vii
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Contents
1 FluorescenceandLabelling:HowtoChooseandWhattoDo. . . . . 1
NataliaFiliandChristopherP.Toseland
2 FluorescentBiosensors:DesignandApplicationto
MotorProteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
SimoneKunzelmann,ClaudiaSolscheid,andMartinR.Webb
3 RapidReactionKineticTechniques. . . . . . . . . . . . . . . . . . . . . . . . 49
ChristopherP.ToselandandMichaelA.Geeves
4 FluorescencetoStudytheATPaseMechanismofMotorProteins. . . 67
ChristopherP.Toseland
5 UseofPyrene-LabelledActintoProbeActin–Myosin
Interactions:KineticandEquilibriumStudies. . . . . . . . . . . . . . . . 87
NancyAdamekandMichaelA.Geeves
6 FluorescentMethodstoStudyTranscriptionInitiationand
TransitionintoElongation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
AishwaryaP.Deshpande,ShemailaSultana,andSmitaS.Patel
7 Single-MoleculeandSingle-ParticleImagingofMolecular
MotorsInVitroandInVivo. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
NataliaFili
8 FluorescenceMethodsintheInvestigationoftheDEAD-Box
HelicaseMechanism. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
AlexandraZ.AndreouandDagmarKlostermeier
9 UseofFluorescentTechniquestoStudytheInVitroMovement
ofMyosins. .. . . . . . . . . .. . . . . . . . . .. . . . . . . . . .. . . . . . . . . .. . 193
ChristopherToepferandJamesR.Sellers
10 FluorescenceTrackingofMotorProteinsInVitro. . . . . . . . . . . . . 211
MarkDeWitt,ThomasSchenkel,andAhmetYildiz
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