Botanisches Institut, Universität Münster, Schloßgarten 3, D-4400, Münster, Federal Republic of Germany.
Planta. 1980 Mar;148(2):168-73. doi: 10.1007/BF00386418.
The non-chloroplastic α-glucan phosphorylase (EC 2.4.1.1) from spinach leaves has been purified to homogeneity as revealed by dodecylsulfate gel electrophoresis. Both purification and separation from the chloroplastic phosphorylase were achieved by chromatography on Sepharose-bound dextrin. The chloroplastic phosphorylase did not bind to Sepharose-dextrin and was removed from the column by washing with buffer, as verified by polyacrylamide gel electrophoresis of the buffer eluate and by chromatography of a preparation from isolated intact chloroplasts. The non-chloroplastic phosphorylase did bind to a high extent to Sepharose-dextrin and could be eluted by a dextrin gradient. Based on dodecylsulfate gel electrophoresis and pyridoxal phosphate determination, a molecular weight of about 90,000 was found for the monomer. Molecular-weight determination by porosity density gradient electrophoresis and gel filtration on Sephadex G-200 suggested that the native enzyme is a dimer, as are other phosphorylases.
菠菜叶片中的非叶绿体 α-葡聚糖磷酸化酶（EC 2.4.1.1）已通过十二烷基硫酸钠凝胶电泳纯化至均质。通过葡聚糖结合的琼脂糖上的色谱分离，可实现从叶绿体磷酸化酶的纯化和分离。叶绿体磷酸化酶不与葡聚糖琼脂糖结合，并用缓冲液洗涤从柱上洗脱，如缓冲液洗脱液的聚丙烯酰胺凝胶电泳和从分离完整叶绿体的制剂的色谱分析所证实的。非叶绿体磷酸化酶高度结合到葡聚糖琼脂糖上，并可通过葡聚糖梯度洗脱。根据十二烷基硫酸钠凝胶电泳和吡哆醛磷酸测定，单体的分子量约为 90,000。通过孔隙率密度梯度电泳和葡聚糖凝胶过滤在 Sephadex G-200 上的分子量测定表明，天然酶是二聚体，与其他磷酸化酶一样。
