酶治疗:聚乙二醇共价附着对-葡萄糖苷酶和-半乳糖糖苷酶生化参数和免疫学决定因素的影响。

Journal of applied biochemistry Pub Date : 1983-08-01
K J Wieder, F F Davis
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引用次数: 0

摘要

聚乙二醇(PEG)与甜杏仁中的-葡萄糖苷酶和绿咖啡豆中的-半乳糖糖苷酶的共价结合导致它们的催化性质的改变和酶上特定决定位点的掩盖。两种酶在PEG连接后,对各自的对硝基苯底物类似物的Km和Vmax值均增加和降低。当PEG与30%的α -半乳糖苷酶ε -氨基结合时,对神经酰胺三己外苷的活性仍为12%,而其将B型红细胞转化为H型红细胞的特异性丧失。然而,它仍然能够从人唾液血型物质b中切割末端半乳糖残基。peg - β -葡萄糖苷酶(38%)不会引起补体固定抗体的产生,也不会与针对天然酶产生的抗体发生反应。两种修饰酶(peg - β -葡萄糖苷酶和peg - α -半乳糖糖苷酶)都失去了抗体和凝集素特异性结合。在与PEG结合后,β -葡萄糖苷酶失去了与魔豆蛋白A-Sepharose结合的能力。针对天然α -半乳糖苷酶的抗体阻断了该酶的活性,但在逐渐高修饰的酶制剂中失去了抑制活性的能力。抗peg - α -半乳糖苷酶的抗血清(53%)不抑制任何α -半乳糖苷酶或peg - α -半乳糖苷酶制剂的酶活性。这些结果表明,聚乙二醇倾向于覆盖凝集素特异性碳水化合物部分和抗原决定因子,这些位点可能在聚乙二醇酶的体内加工过程中保持隐性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Enzyme therapy: II. Effect of covalent attachment of polyethylene glycol on biochemical parameters and immunological determinants of beta-glucosidase and alpha-galactosidase.

The covalent attachment of polyethylene glycol (PEG) to beta-glucosidase from sweet almonds and alpha-galactosidase from green coffee beans results in alterations of their catalytic properties and masking of specific determinant sites on the enzymes. Both enzymes have increased Km and decreased Vmax values against their respective p-nitrophenyl substrate analogs after PEG attachment. When PEG is attached to 30% of alpha-galactosidase epsilon-amino groups, 12% activity remains against ceramide trihexoside, while its ability to convert type B erythrocytes to type H specificity is lost. However, it still is able to cleave terminal galactose residues from human saliva blood group substance B. PEG-beta-glucosidase (38%) did not elicit the production of complement-fixing antibodies, nor did it react with antibodies produced against the native enzyme. Antibody and lectin-specific binding were lost from both modified enzymes (PEG-beta-glucosidase and PEG-alpha-galactosidase). After conjugation with PEG, beta-glucosidase lost its ability to bind to concanavalin A-Sepharose. Antibodies directed against native alpha-galactosidase blocked its enzyme activity, but lost their ability to inhibit activity in progressively higher modified preparations of the enzyme. Antisera against PEG-alpha-galactosidase (53%) did not inhibit enzyme activity in any alpha-galactosidase or PEG-alpha-galactosidase preparation. These results indicate that PEG tends to cover lectin-specific carbohydrate moieties and antigenic determinants and that these sites probably remain cryptic during in vivo processing of PEG-enzymes.

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