A comparative study on alpha-glucan phosphorylases from plant and animal: interrelationship between the polysaccharide and pyridoxal phosphate binding sites by affinity electrophoresis.
A comparative study on alpha-glucan phosphorylases from plant and animal: interrelationship between the polysaccharide and pyridoxal phosphate binding sites by affinity electrophoresis.
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植物和动物α-葡聚糖磷酸化酶的比较研究：通过亲和电泳研究多糖和磷酸吡哆醛结合位点之间的相互关系。
DOI：
10.1021/bi00552a001
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发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
T. Fukui
中科院分区：
文献类型：
--
作者：
S. Shimomura;T. Fukui
Shoji Shimomura和Toshio福井 * 摘要：磷酸化酶（EC 2.4. 1.1)通过比较完整酶和在辅因子位点修饰的酶对葡聚糖的亲和力，研究了从马铃薯块茎和兔骨骼肌中提取的葡聚糖。蛋白质-糖复合物的解离常数通过在聚丙烯酰胺凝胶中的亲和电泳测定。该程序允许通过电泳迁移率的降低来评估蛋白质与凝胶基质中存在的配体相互作用的程度。通过用在5 '位修饰的辅因子（吡哆醛5'-磷酸）类似物重构脱辅基酶来制备修饰的磷酸-淀粉酶。兔肌磷酸化酶B对糖原的亲和力几乎不受修饰的影响。相比之下，马铃薯磷酸化酶对支链淀粉和麦芽五糖的亲和力受到以下特征性影响：
Shoji Shimomura and Toshio Fukui* abstract: The interrelationship between the cofactor and al, 4-glucan binding sites in phosphorylases (EC 2.4. 1.1) from potato tubers and rabbit skeletal muscle has been investigated by comparing the affinities of the intact enzyme and the en-zyme modified at the cofactor site for the glucan. The dissociation constants of the protein-saccharide complexes were determined by affinity electrophoresis in polyacrylamide gel. This procedure allows one to evaluate the extent to which a protein interacts with a ligand present in the gel matrix by the decrease in electrophoretic mobility. The modified phospho-rylases were prepared by reconstitution of the apoenzymes with the cofactor (pyridoxal 5'-phosphate) analogues modified at the 5'position. The affinity of rabbit muscle phosphorylase b for glycogen was little affected by the modifications. By contrast, the affinity of potato phosphorylase for amylopectin and maltopentaose was characteristically affected according