Sequential Extraction Results in Improved Proteome Profiling of Medicinal Plant Pinellia ternata Tubers, Which Contain Large Amounts of High-Abundance Proteins 英文参考文献.docVIP

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Sequential Extraction Results in Improved Proteome Profiling of Medicinal Plant Pinellia ternata Tubers, Which Contain Large Amounts of High-Abundance Proteins 英文参考文献.doc

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Sequential Extraction Results in Improved Proteome Profiling of Medicinal Plant Pinellia ternata Tubers, Which Contain Large Amounts of High-Abundance Proteins 英文参考文献

SequentialExtractionResultsinImprovedProteome ProfilingofMedicinalPlantPinelliaternataTubers, WhichContainLargeAmountsofHigh-Abundance Proteins XiaoLinWu.,ErHuiXiong.,SuFangAn,FangPingGong,WeiWang* KeyLaboratoryofPhysiologicalEcologyandGeneticImprovementofFoodCropsinHenanProvince,CollegeofLifeScience,HenanAgriculturalUniversity,Zhengzhou, China Abstract Pinelliaternatatuberisoneofthewell-knownChinesetraditionalmedicines.Inordertounderstandthepharmacological properties of tuber proteins, it is necessary to perform proteome analysis of P. ternata tubers. However, a few high- abundanceproteins(HAPs),mainlymannose-bindinglectin(agglutinin),existinaggregatesofvarioussizesinthetubers andseriouslyinterferewithproteomeprofilingbytwo-dimensionalelectrophoresis(2-DE).Therefore,selectivedepletionof theseHAPsisaprerequisiteforenhancedproteomeanalysisofP.ternatatubers.Basedondifferentialproteinsolubility,we developedanovelprotocolinvolvingtwosequentialextractionsfordepletionofsomeHAPsandprefractionationoftuber proteins prior to 2-DE. The first extraction using 10% acetic acid selectively extracted acid-soluble HAPs and the second extractionusingtheSDS-containingbufferextractedremainingacid-insolubleproteins.Afterapplicationoftheprotocol,2- DE profiles of P. ternata tuber proteins were greatly improved and more protein spots were detected, especially low- abundanceproteins.Moreover,thesubunitcompositionofP.ternatalectinwasanalyzedbyelectrophoresis.Nativelectin consistsoftwohydrogen-bondedsubunits(11kDaand25kDa)andthe11kDasubunitwasaglycoprotein.Subsequently, majorHAPsinthetuberswereanalyzedbymassspectrometry,withnineproteinspotsbeingidentifiedaslectinisoforms. Themethodologywaseasytoperformandrequirednospecializedapparatus.Itwouldbeusefulforproteomeanalysisof othertuberplantsofAraceae. Citation:WuX,XiongE,AnS,GongF,WangW(2012)SequentialExtractionResultsinImprovedProteomeProfilingofMedicinalPlantPinelliaternataTubers, WhichContainLargeAmountsofHigh-AbundanceProteins.PLoSONE7(11)