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盘基网柄菌无菌培养的变形虫中UDP-葡萄糖焦磷酸化酶的积累。

Accumulation of UDP-glucose pyrophosphorylase by axenically grown amoebae of Dictyostelium discoideum.

作者信息

Hames B D, Hodson B A

出版信息

Biochem J. 1979 Jan 1;177(1):21-8. doi: 10.1042/bj1770021.

Abstract

Amoebae of the slime mould Dictyostelium discoideum AX2 possess only low UDP-glucose pyrophosphorylase activity when grown on autoclaved Klebsiella aerogenes (approx. 30 units/mg of protein), but accumulate the enzyme to approx. 150-200 units/mg of protein during vegetative growth in axenic medium. The vegetative accumulation of UDP-glucose pyrophosphorylase by axenically grown cells is prevented if autoclaved K. aerogenes are included in the axenic medium, suggesting the absence of a specific inducer. Affinity chromatography using anti-(UDP-glucose pyrophosphorylase) antibody and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicate that the enzyme accumulated during axenic growth and that normally accumulated during development are immunologically cross-reactive and that both are composed of two subunits with mol.wts. 55,600 and 57,500 present in approximately equal amounts in the active enzyme.

摘要

当黏菌盘基网柄菌AX2的变形虫在经高压灭菌的产气克雷伯菌上生长时,其UDP - 葡萄糖焦磷酸化酶活性很低(约30单位/毫克蛋白质),但在无菌培养基中营养生长期间,该酶会积累至约150 - 200单位/毫克蛋白质。如果在无菌培养基中加入经高压灭菌的产气克雷伯菌,无菌培养的细胞中UDP - 葡萄糖焦磷酸化酶的营养积累就会受到抑制,这表明不存在特定的诱导物。使用抗(UDP - 葡萄糖焦磷酸化酶)抗体的亲和层析和十二烷基硫酸钠/聚丙烯酰胺凝胶电泳表明,在无菌生长期间积累的酶以及正常发育期间积累的酶在免疫上具有交叉反应性,并且两者均由两个亚基组成,活性酶中分子量分别为55,600和57,500的亚基含量大致相等。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76b4/1186336/ec9b1bcbd548/biochemj00471-0035-a.jpg

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