Alkaline phosphatase of sea urchin embryos: Chromatographic and electrophoretic characterization☆
[摘要] Achangeinthemolecularformofalkalinephosphataseinseaurchinembryosaccompaniesthemarkedincreaseinactivitythatoccursatgastrulation.Onthebasisofchromatographicandelectrophoreticanalyses,twomajorclassesofalkalinephosphataseareidentified:earlyenzyme,fromunfertilizedeggstomesenchymeblastula,characterizedbyamajorpeakofactivity,withaKavof0.123onSephadexG-200columns,elutionfromDEAE-Sephadexcolumnsby0.5MNaCl,andamigrationvalueof0.51(relativetobromophenolblue)afterelectrophoresisin7.5%polyacrylamidegels;lateenzyme,fromgastrulatoplutei,characterizedbyaKavof0.137,elutionfromDEAE-Sephadexby0.55–0.75MNaCl,andamigrationvalueof0.56.Bychromatographicandelectrophoreticcriteriatheearlyenzymeappearstohaveaslightlygreatermolecularvolume,lowernetnegativecharge,andmoreheterogeneouscompositionthanthelateenzyme.BothenzymepreparationsweremaximallyactiveatapH9.4–9.5.Enzymefromallstagesappearstobepredominantlyassociatedwithcellmembranes.Extractingtheenzymebytreatmentwithn-butanol,precipitatingtheenzymefromthedialyzedaqueousphasewithethylalcohol,andchromatographingthealcoholpreparationoncolumnsofsievingandanion-exchangingmediaresultedinasubstantialpurificationoftheenzymefromallstages.
[发布日期] [发布机构]
[效力级别] [学科分类] 生物科学（综合）
[关键词] [时效性]
