Calibration and standardisation of synchrotron radiation and conventional circular dichroism spectrometers. Part 2 Factors affecting magnitude and wavelength英文文献资料.docVIP

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Calibration and standardisation of synchrotron radiation and conventional circular dichroism spectrometers. Part 2 Factors affecting magnitude and wavelength英文文献资料.doc

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Calibration and standardisation of synchrotron radiation and conventional circular dichroism spectrometers. Part 2 Factors affecting magnitude and wavelength英文文献资料

Spectroscopy19(2005)43–51 IOSPress 43 Calibrationandstandardisation ofsynchrotronradiationandconventional circulardichroismspectrometers.Part2: Factorsaffectingmagnitudeandwavelength AndrewJ.Miles ,FrankWien a a,JonathanG.Leesa,?andB.A.Wallacea,b,?? aDepartmentofCrystallography,BirkbeckCollege,UniversityofLondon,LondonWC1E7HX, UnitedKingdom bCentreforProteinandMembraneStructureandDynamics,DaresburyLaboratory, WarringtonWA44AD,UnitedKingdom Abstract.Circulardichroism(CD)spectroscopy isanimportanttoolinstructuralbiology, especiallyforproteinsecondary structureanalyses.Synchrotronradiationcirculardichroism(SRCD)spectroscopyisamodi?edversionofthetechniquethat uses theintense light froma synchrotron source to enable the collection of data tomuch lower wavelengths thanpossible on conventional circular dichroism (cCD) instruments. There isa need for standardization of calibration methods amongst andbetweencCDandSRCDinstrumentstoensureconsistencyandtheabilitytousecommonreferencedatabasesforem- piricalsecondary structuralanalyses.Inapreviousstudy(Spectroscopy 17(2003),653–661), wecomparedopticalrotation measurementsonseveralcCDandSRCDinstruments,whilstholdingconstantotherexperimentalfactors.Inthisstudy,other experimentalparameterswhichcontributetothespectralmagnitude,suchascellpathlengthandproteinconcentrationdeter- minations, are examined. In addition, the extent of wavelength calibration variations between instruments and their effects onsecondarystructurecalculationshavebeenexamined.Hence,thispaperprovidesadditionalpracticalguidancefor“good practice”inthemeasurementofCDdata. 1. Introduction Circulardichroism(CD)spectroscopyisalong-establishedtechniqueforexaminingthesecondary structure of proteins and for monitoring protein interactions and folding pathways. It is gaining ac- ceptanceasatechniqueapprovedforthecharacterizationofmacromoleculesforpharmacologicaluse. Synchrotronradiation-basedcirculardichroism(SRCD)spectroscopymadeitsdebutmorethan20year