Characterization of concanavalin A precipitated proteins from early mouse embryos: A 2-dimensional gel electrophoresis study☆

[摘要] ConcanavalinA(ConA),aplantlectinwhichbindstomannose-containingcomponents,hasbeenusedtoprecipitateaspecificclassofproteinssynthesizedduringpreimplantationmouseembryogenesis.TotalcellularproteinwasextractedfromlabeledembryoswithNonidetP40,reactedwithConA,andthenexposedtorabbitantiserumdirectedagainstConA.ImmunecomplexeswereprecipitatedwithproteinAfromStaphylococcusaureus.Afterwashing,ConA-boundproteinswereelutedwithmannosidewhichreleasedbetween3and10%ofthetotalradioactivityfromunfertilizedandfertilizedeggs,2-celland8-to16-cellembryos,andlateblastocysts.Incontrolexperiments,inwhichmannosidewasaddedpriortotheConA,only0.1–0.5%ofthetotalradioactivitywaseluted,indicatinglittlereleaseofproteinsnonspecificallyboundtotheproteinA-anti-ConA-ConAcomplex.Whentheseprecipitateswereexaminedbytwo-dimensionalgelelectrophoresis,approximately30–70peptidesinthedefinedperiodsofsynthesisweredetectedinautoradiograms.Toprecipitatecellsurfaceproteins,intactembryoswereincubatedinConAandanti-ConApriortoextractionwithNonidetP40.Lessthan1.0%ofthetotalradioactivitywaselutedwithmannoside.Acomplexofthreepairsofpeptideswaspresentin2-celland8-to16-cellembryos.Thequalitativechangesintwo-dimensionalgelpatternsofConA-bindingproteinshasprovidedaprogramofsynthesisoccurringduringnormaldevelopmentforaspecificclassofproteins.

[学科分类] 生物科学（综合）