Fluorescent Methods for Molecular Motors PDF
Preview Fluorescent Methods for Molecular Motors
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 ThiSisaFMBlankPage 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 Thisworkissubjecttocopyright.AllrightsarereservedbythePublisher,whetherthewholeorpart of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation,broadcasting,reproductiononmicrofilmsorinanyotherphysicalway,andtransmissionor informationstorageandretrieval,electronicadaptation,computersoftware,orbysimilarordissimilar methodologynowknownorhereafterdeveloped.Exemptedfromthislegalreservationarebriefexcerpts inconnectionwithreviewsorscholarlyanalysisormaterialsuppliedspecificallyforthepurposeofbeing enteredandexecutedonacomputersystem,forexclusiveusebythepurchaserofthework.Duplication ofthispublicationorpartsthereofispermittedonlyundertheprovisionsoftheCopyrightLawofthe Publisher’s location, in its current version, and permission for use must always be obtained from Springer.PermissionsforusemaybeobtainedthroughRightsLinkattheCopyrightClearanceCenter. ViolationsareliabletoprosecutionundertherespectiveCopyrightLaw. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publicationdoesnotimply,evenintheabsenceofaspecificstatement,thatsuchnamesareexempt fromtherelevantprotectivelawsandregulationsandthereforefreeforgeneraluse. While the advice and information in this book are believed to be true and accurate at the date of publication,neithertheauthorsnortheeditorsnorthepublishercanacceptanylegalresponsibilityfor anyerrorsoromissionsthatmaybemade.Thepublishermakesnowarranty,expressorimplied,with respecttothematerialcontainedherein. Printedonacid-freepaper SpringerispartofSpringerScience+BusinessMedia(www.springer.com) For Thalia...You remind us of how much we can learn every day. ThiSisaFMBlankPage 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 ThiSisaFMBlankPage 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 ix
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