Proteomic characterisation of endoplasmic reticulum-derived protein bodies in tobacco leaves 英文参考文献.docVIP
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Proteomic characterisation of endoplasmic reticulum-derived protein bodies in tobacco leaves 英文参考文献
Josephetal.BMCPlantBiology2012,12:36
/1471-2229/12/36
RESEARCH ARTICLE
OpenAccess
Proteomiccharacterisationofendoplasmic
reticulum-derivedproteinbodiesintobacco
leaves
MinuJoseph2,MDolorsLudevid2,MargaritaTorrent2,ValérieRofidal1,MarcTauzin1,MichelRossignol1and
Jean-BenoitPeltier1*
Abstract
Background:TheN-terminalproline-richdomain(Zera)ofthemaizestorageproteing-zein,isabletoinducethe
formationofendoplasmicreticulum(ER)-derivedproteinbodies(PBs)whenfusedtoproteinsofinterest.This
encapsulationenablesarecombinantfusedproteintoescapefromdegradationandfacilitatesitsrecoveryfrom
plantbiomassbygradientpurification.TheaimofthepresentworkwastoevaluateifinducedPBsencapsulate
additionalproteinsjointlywiththerecombinantprotein.TheexhaustiveanalysisofproteincompositionofPBsis
expectedtofacilitateabetterunderstandingofPBformationandtheoptimizationofrecombinantprotein
purificationapproachesfromtheseorganelles.
Results:WeanalysedtheproteomeofPBsinducedinNicotianabenthamianaleavesbytransienttransformation
withZerafusedtoafluorescentmarkerprotein(DsRed).IntactPBswiththeirsurroundingER-membranewere
isolatedoniodixanolbaseddensitygradientsandtheirintegrityverifiedbyconfocalandelectronmicroscopy.SDS-
PAGEanalysisofisolatedPBsshowedthatZera-DsRedaccountedforaround85%ofPBproteinsintermof
abundance.DifferentialextractionofPBswasperformedforin-depthanalysisoftheirproteomeandstructure.
BesidesZera-DsRed,195additionalproteinswereidentifiedincludingabroadrangeofproteinsresidentor
traffickingthroughtheERandrecruitedwithintheZera-DsRedpolymer.
Conclusions:ThisstudyindicatesthatZera-proteinfusionisstillthemajorproteincomponentofthenewformed
organelleintobaccoleaves.TheanalysisalsorevealsthepresenceofanunexpecteddiversityofproteinsinPBs
derivedfromboththeinsolubleZera-DsRedpolymerformation,includingER-residentandsecretoryproteins,anda
secretorystressresponseinducedmostlikelybytherecombinantproteinoverloading.KnowledgeofPBsprotein
compositionislikelytobeusefultooptimizedownstreampurificationof
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