Anchoring Secreted Proteins in Endoplasmic Reticulum by Plant Oleosin The Example of Vitamin B12 Cellular Sequestration by Transcobalamin 英文参考文献.docVIP
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Anchoring Secreted Proteins in Endoplasmic Reticulum by Plant Oleosin The Example of Vitamin B12 Cellular Sequestration by Transcobalamin 英文参考文献
AnchoringSecretedProteinsinEndoplasmicReticulum
byPlantOleosin:TheExampleofVitaminB12Cellular
SequestrationbyTranscobalamin
LaurentPons1,2.,Shyue-FangBattaglia-Hsu1.,CarlosEnriqueOrozco-Barrios3.,SandrineOrtiou1.
CelineChery1.,Jean-MarcAlberto1,MarthaLigiaArango-Rodriguez3,DominiqueDumas4 ,Daniel
Martinez-Fong3,Jean-NoelFreund5,Jean-LouisGueant1*
,
1Inserm U954, Faculte′ de Me′decine, Nancy-Universite′, Vandoeuvre-les-Nancy, France, 2Department of Pediatrics, Duke University Medical Center, Durham, North
Carolina,UnitedStatesofAmerica,3DepartmentofPhysiology,BiophysicsandNeuroscience,CINVESTAV,MexicoD.F.,Mexico,4PlateformeIBISAd’imagerieetbiologie
cellulaire(PTIBC),UMRCNRS7561,FRCNRS/Inserm3209,Faculte′ deMe′decine,Nancy-Universite′,Vandoeuvre-les-Nancy,France,5INSERM,U682,Strasbourg,France
Abstract
Background:Oleosinisaplantproteinlocalizedtolipiddropletsandendoplasmicreticulumofplantcells.Ourideawasto
useittotargetfunctionalsecretoryproteinsofinteresttothecytosolicsideoftheendoplasmicreticulumofmammalian
cells,throughexpressingoleosin-containingchimeras.Wehavedesignedthisapproachtocreatecellularmodelsdeficient
in vitamin B12 (cobalamin) because of the known problematics associated to the obtainment of effective vitamin B12
deficientcellmodels.Thiswasachievedbytheoverexpressionoftranscobalamininsidecellsthroughanchoringtooleosin.
Methodology:chimerageneconstructsincludingtranscobalamin-oleosin(TC-O),greenfluorescentprotein-transcobalamin-
oleosin(GFP-TC-O)andoleosin-transcobalamin(O-TC)wereinsertedintopAcSG2andpCDNA3vectorsforexpressioninsf9
insect cells, Caco2 (colon carcinoma), NIE-115 (mouse neuroblastoma), HEK (human embryonic kidney), COS-7 (Green
Monkey SV40-transfected kidney fibroblasts) and CHO (Chinese hamster ovary cells). The subcellular localization, the
changesinvitaminB12bindingactivityandthemetabolicconsequenceswereinvestigatedinbothCaco2andNIE-115cells.
Principal findings: vitamin B12 binding was dramatically higher in TC-O than that in O-TC
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