Some Properties of a Macromolecular Conjugate of Lysozyme Prepared by Modification with a Monomethoxypolyethylene Glycol Derivative
Some Properties of a Macromolecular Conjugate of Lysozyme Prepared by Modification with a Monomethoxypolyethylene Glycol Derivative
复制标题
单甲氧基聚乙二醇衍生物修饰溶菌酶大分子结合物的一些性质
DOI：
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
N. Yamasaki
中科院分区：
文献类型：
--
作者：
Y. Nodake;N. Yamasaki
用单甲氧基聚乙二醇的琥珀酰酯衍生物（mPEG-COONSu）修饰鸡卵清溶菌酶，并对所得偶联物（mPEG-溶菌酶）的性质进行了研究。用mPEG-COONSu修饰溶菌酶制备偶联物，用CM-Toyopolymer 650 M和SephadexG-75柱纯化。分析数据表明，在缀合物中，1.05摩尔的平均分子量为5，000的mPEG共价连接至溶菌酶分子。结合物的胰蛋白酶肽分析表明，溶菌酶中的Lys 33是mPEG-COONSu主要修饰的残基。共价连接的mPEG-衍生物的氨基大大增加了溶菌酶的热稳定性，而没有任何构象的蛋白质分子的变化。mPEG-溶菌酶对乙二醇甲壳素仍保持完全的酶活性，但在中性介质中对藤黄微球菌细胞的裂解活性为天然溶菌酶的75%，其最适pH为pH5.0。经mPEG-COONSu修饰后，溶菌酶与BALB/c小鼠抗溶菌酶抗体或人淋巴细胞抗溶菌酶抗体的反应性降低。根据这些发现，表明mPEG-COONSu可以有利地用于溶菌酶的蛋白质剪裁。
Hen egg-white lysozyme was modified with a succinyl ester derivative of monomethoxypolyethylene glycol (mPEG-COONSu), and some properties of the resulting conjugate (mPEG-lysozyme) were studied. The conjugate was prepared by modification of lysozyme with mPEG-COONSu and purified with use of columns of CM-Toyopearl 650M and Sephadex G-75. Analytical data indicated that in the conjugate, 1.05 moles of mPEG with an average molecular weight of 5,000 were covalently attached to the lysozyme molecule. Tryptic peptide analysis of the conjugate showed that Lys 33 in lysozyme is the residue mainly modified with mPEG-COONSu. Covalent attachment of the mPEG-derivative to amino groups greatly increased the thermostability of lysozyme without any conformational change of the protein molecule. mPEG-lysozyme retained full enzyme activity for glycol chitin, but lytic activity for Micrococcus luteus cells in neutral media was 75% of that of native lysozyme and its optimal pH was at pH 5.0. It was also found that the reactivity of lysozyme with anti-lysozyme antibody from BALB/c mice or human lymphocytes was decreased by modification with mPEG-COONSu. From these findings, it was suggested that mPEG-COONSu can be advantageously used for protein tailoring of lysozyme.
影响因子：
158.5
作者：
HERSHFIELD, MS;BUCKLEY, RH;ABUCHOWSKI, A
通讯作者：
ABUCHOWSKI, A
DOI：
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发表时间：
1981
期刊：
International archives of allergy and applied immunology
影响因子：
--
作者：
Wie,SI;Wie,CW;Lee,WY;Filion,LG;Sehon,AH;Akerblom,E
通讯作者：
Akerblom,E