A Hydrazine Coupled Cycling Assay Validates the Decrease in Redox Ratio under Starvation in Drosophila 英文参考文献.docVIP
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A Hydrazine Coupled Cycling Assay Validates the Decrease in Redox Ratio under Starvation in Drosophila 英文参考文献
AHydrazineCoupledCyclingAssayValidatesthe
DecreaseinRedoxRatiounderStarvationinDrosophila
Chen-TsehZhu*,DavidM.Rand
DepartmentofEcologyandEvolutionBiology,BrownUniversity,Providence,RhodeIsland,UnitedStatesofAmerica
Abstract
AcommonlyusedenzymaticrecyclingassayforpyridinenucleotideshasbeenadaptedtodirectlymeasuretheNAD+/NADH
redoxratioinDrosophilamelanogaster.ThismethodisalsosuitableforquantificationofNADP+andNADPH.Theadditionof
a coupling reaction removing acetaldehyde produced from the alcohol dehydrogenase (ADH) reaction was shown to
improvethelinearityofNAD(H)assay.Theadvantagesofthisassaymethodarethatitallowsthedeterminationofboth
+
NAD andNADHsimultaneouslywhilekeepingenzymaticdegradationofpyridinenucleotidesminimalandalsoachieving
bettersensitivity.ThismethodwasusedtodeterminetheredoxratioofD.melanogasterandvalidatedsubstantialdecrease
ofredoxratioduringstarvation.
Citation:ZhuC-T,RandDM(2012)AHydrazineCoupledCyclingAssayValidatestheDecreaseinRedoxRatiounderStarvationinDrosophila.PLoSONE7(10):
e47584.doi:10.1371/journal.pone.0047584
Editor:LisaCarlsonLyons,FloridaStateUniversity,UnitedStatesofAmerica
ReceivedMay10,2012;AcceptedSeptember13,2012;PublishedOctober17,2012
Copyright: ? 2012 Zhu, Rand. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits
unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited.
Funding:ThisstudywassupportedbygrantsfromtheNationalInstitutesofHealth:1RO1AG027849and2RO1GM067862,bothtoD.M.R.Thefundershadnorole
instudydesign,datacollectionandanalysis,decisiontopublish,orpreparationofthemanuscript.
CompetingInterests:Theauthorshavedeclaredthatnocompetinginterestsexist.
*E-mail:ctzhu@
Introduction
Since the assay will not distinguish between reduced and
+
oxidized pyridine nucleotides, to measure NAD /NADH redox
The levels of pyridine nucleotides and their redox ratios,
NAD+/NADH and NADP+/NADPH, are important biological
signatur
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