Purification of cortical granules from unfertilized sea urchin egg homogenates by zonal centrifugation☆

[摘要] Corticalgranuleswerepurifiedfromhomogenatesofunfertilizedseaurchin,Strongylocentrotuspurpurataus,eggsbyrate-zonalcentriguation.Purificationwasevaluatedbyintegratedbiochemicalandmorphological(electronmicroscopic)analysis.Theenzymeβ-1,3-glucanasewasemployedasthebiochemicalmarkerforcorticalgranules.Corticalgranulepreparationsobtainedinsucrosegradientsconstructedin0.5MKClcontainedacidphosphataseactivityandwereheavilycontaminatedbyyolkplatelets.Highlypurifiedcorticalgranulesisolatedinsucrosegradientsconstructedin0.5MKClwith10−3MEDTAatpH7.0werealmostcompletelyfreeofcontaminationbyyolkandothercytoplasmicparticles,andcontainedonlytracelevelsofacidphosphataseactivity.Thepurifiedcorticalgranulescontained40–45%ofthetotalβ-1,3-glucanaseactivityintheeggs,and6.7±1.9%ofthetotaleggprotein.Theisolatedcorticalgranulesretainedanotherknownchemicalconstituent,sulfatedacidmucopolysaccharides.

[学科分类] 生物科学（综合）