Facile Method of Protein PEGylation by a Mono-Ion Complex
Facile Method of Protein PEGylation by a Mono-Ion Complex
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DOI：
10.1021/acsomega.7b00462
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发表时间：
2017-05
期刊：
影响因子：
4.1
通讯作者：
S. Asayama;Kana Nagashima;H. Kawakami
中科院分区：
文献类型：
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作者：
S. Asayama;Kana Nagashima;H. Kawakami
合成了用于蛋白质非共价聚乙二醇化反应的二乙氨基乙基末端修饰聚乙二醇酯(DEAE-PEG)。生成的DEAE-PEG和过氧化氢酶形成离子复合体，即蛋白质单离子复合体(MIC)。天然聚丙烯酰胺凝胶电泳法和凝胶过滤层析法证实了MIC蛋白的形成。得到的过氧化氢酶MIC保留了过氧化氢酶的活性，通过克拉克型氧电极监测O2浓度证实了这一点，尽管MIC形成了。蛋白MIC的过氧化氢酶活性在蛋白酶、胰酶或10%胎牛血清的存在下得到保护。此外，与过氧化氢酶-聚乙二醇偶联物(共价聚乙二醇化)相比，过氧化氢酶MIC的圆二色谱测量变化较小，这表明蛋白质构象的影响较小。因此，MIC的形成被认为是一种简便的蛋白质聚乙二醇化方法。
Diethylaminoethyl end-modified poly(ethylene glycol) (DEAE-PEG) has been synthesized for the noncovalent PEGylation of proteins. The resulting DEAE-PEG and catalase formed an ion complex, that is, a protein mono-ion complex (MIC). The formation of the protein MIC was confirmed by native poly(acrylamide) gel electrophoresis and gel-filtration chromatography. The resulting catalase MIC preserved the catalase activity, confirmed by monitoring the O2 concentration with a Clark-type oxygen electrode, in spite of MIC formation. The catalase activity of the protein MIC was protected in the presence of a protease, trypsin, or 10% fetal bovine serum. Furthermore, less change in the circular dichroism measurements of the catalase MIC was observed as compared to those of a catalase–PEG conjugate (covalent PEGylation), suggesting less influence of the protein conformation. Consequently, the formation of the MIC is considered to be a facile method of protein PEGylation.