Kinetic Studies on Allosteric Properties of Honeybee alpha-Glucosidase

蜜蜂α-葡萄糖苷酶变构特性的动力学研究

• 批准号: 02660074
• 负责人: CHIBA Seiya
• 金额: $ 1.28万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02660074/
• 关键词: Honeybee; alpha-Glucosidase; Allosteric effect; Cooperativity; αーグルコンダ-ゼ

(1)Three kinds of alpha-glucosidases(HbG-I, HbG-II and HbG-III)were purified from honeybee. In addition, an alpha-glucosidase was also purified from honey.(2)HbG-I, II and III were immunologically confirmed to be distributed in ventriculus, hemolymph(and also ventriculus)and hypopharyngeal gland, respectively. The alpha-glucosidase from honey was identified to be the same enzyme as HbG-III.(3)HbG-I and II showed allosteric behaviors. These two enzymes were regarded as so-called hysteretic enzyme which showed a relatively slow conformational change in second or minute order.(4)The subsites affinities in active site of HbG-I, II and III were evaluated. Subsite 1, 2 and 3 having positive values in three enzymes were considered to be effective for binding of substrate to active site.(5)Honeybee alpha-glucosidase, which is the same enzyme as HbG-III, was considered to be secreted into honeydew from hypopharyngeal gland.

（1）从蜜蜂中分离纯化了三种α-葡萄糖苷酶（HbG-I、HbG-II和HbG-III）。此外，还从蜂蜜中纯化了α-葡糖苷酶。（2）免疫组化证实，HbG-I、HbG-II、HbG-III分别分布于胃、血淋巴和下咽腺。蜂蜜中的α-葡萄糖苷酶被鉴定为与HbG-III相同的酶。（3）HbG-I和HbG-II具有变构行为。这两种酶被认为是所谓的滞后酶，表现出相对缓慢的构象变化在秒或分钟的顺序。（4）计算HbG-I、II、III活性位点的亲和力。在三种酶中具有正值的亚位点1、2和3被认为对底物与活性位点的结合有效。（5）蜜蜂α-葡萄糖苷酶与HbG-III是同一种酶，被认为是从下咽腺分泌到蜜露中的。

项目成果

Sumio Kitahata: "Mechanism of Maltal Hydration Catalyzed by beta-Amylase : Role of Protein Structure in Controlling the Steric Outcome of Reaction Catalyzed by a Glycosylase" Biochemistry. 30. 6769-6775 (1991)

Sumio Kitahata：“β-淀粉酶催化的麦芽糖水合机制：蛋白质结构在控制糖基酶催化反应的空间结果中的作用”生物化学。

K.Kawagishi: "Isolation and Characterization of A Lectin from Grifola frondosa Fruiting Bodies." Biochim.Biophys.Acta,. 1034. 247-252 (1990)

K.Kawagishi：“灰树花子实体中凝集素的分离和表征”。

Hirokazu Kawagishi: "A Lectin from the Mushroom aurirella" Agric.Biol.Chem.55. 2485-2489 (1991)

Hirokazu Kawagishi：“来自蘑菇 aurirella 的凝集素”Agric.Biol.Chem.55。

Hirokazu Kawagishi: "Isolation and Characterization of A lection from Grifola frondosa Fruiting Bodies" Biochim. Biophys. Acta. 1034. 247-252 (1990)

Hirokazu Kawagishi：“灰树花子实体的分离和表征”Biochim。

A.Kita: "Substrate Specificity and Subsite Affinifies of Crystalline αーglucosidase from Asopergillus niger" Agric.Biol.Chem.55. 2327-2335 (1991)

A.Kita：“黑曲霉结晶 α-葡萄糖苷酶的底物特异性和亚位点亲和力”Agric.Biol.Chem.55 (1991)。