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首页> 外文期刊>Journal of bacteriology >Purification and Properties of the Periplasmic Glucose-Binding Protein of Pseudomonas aeruginosa
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Purification and Properties of the Periplasmic Glucose-Binding Protein of Pseudomonas aeruginosa

机译：铜绿假单胞菌周质葡萄糖结合蛋白的纯化和性质

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摘要

A glucose-binding glycoprotein (GBP) from the periplasm of Pseudomonas aeruginosa was purified to homogeneity as judged by polyacrylamide gel electrophoresis, molecular sieve chromatography, and double-diffusion gel precipitation. It had an average molecular weight of 44,500 and an isoelectric point of 4.7. One mole of glucose was bound per mole of GBP with a dissociation constant of 0.35 μM. The binding of radioactive glucose by GBP was not significantly inhibited by 10-fold-higher concentrations of other carbohydrates; however, a number of related compounds were found to compete at 100-fold-higher concentrations. Amino acid analyses revealed predominant amounts of alanine, glutamate, and glycine and a low content of sulfur-containing amino acids. The carbohydrate moiety of GBP, comprising nearly 16% of the total weight, contained galactosamine, glucosamine, fucose, galactose, glucose, and mannose. A GBP-deficient mutant, strain MB723, was found to be defective in both membrane transport and glucose chemotaxis. Strain MB724, a revertant to GBP-positive phenotype, simultaneously recovered normal levels of both membrane functions.
机译：根据聚丙烯酰胺凝胶电泳，分子筛色谱和双扩散凝胶沉淀法，将铜绿假单胞菌周质中的葡萄糖结合糖蛋白（GBP）纯化至均一。其平均分子量为44,500，等电点为4.7。每摩尔GBP结合1摩尔葡萄糖，其解离常数为0.35μM。 GBP对放射性葡萄糖的结合没有被其他碳水化合物的10倍高浓度显着抑制。但是，发现许多相关化合物的竞争浓度要高100倍。氨基酸分析表明，丙氨酸，谷氨酸和甘氨酸占主导地位，而含硫氨基酸含量低。 GBP的碳水化合物部分占总重量的近16％，其中包含半乳糖胺，氨基葡萄糖，岩藻糖，半乳糖，葡萄糖和甘露糖。一个GBP缺陷突变体菌株MB723被发现在膜转运和葡萄糖趋化性方面均存在缺陷。 MB724菌株是GBP阳性表型的回复株，可同时恢复两种膜功能的正常水平。

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