标题：Development of a method for the efficient release of N-glycans from glycoproteins generating native deglycosylated proteins
作者机构：[Wang, S] State Key Laboratory of Microbial Technology, Shandong University, Jinan, Shandong 250100, China;[ Ma, C] State Key Laboratory of Microbial 更多
通讯作者地址：[Qi, QS]Shandong Univ, Sch Life Sci, State Key Lab Microbial Technol, Shanda Nanlu 27, Jinan 250100, Shandong, Peoples R China.
来源：Enzyme and Microbial Technology
关键词：Pnglp-ΔHl;Dithiothreitol;Deglycosylation;Refolding;Native deglycosylated proteins
摘要：In many cases, it is desirable to maintain the native status of the target glycoproteins when they are deglycosylated. However, most conventional deglycosylation process often causes the irreversible denat-uration of the target glycoproteins. In the present study, we developed a deglycosylation method that could obtain the native deglycosylated proteins employing Pnglp-ΔHl, which was confirmed to tolerate high concentration of dithiothreitol (DTT). To prove this process, ribonuclease B (RNase B) and Yeast carboxypeptidase (CPY) were employed as the targeting glycoproteins. Our results confirmed that both of them could be completely deglycosylated in the presence of high concentration DTT and could be refolded when DTT was removed. The circular dichroism spectroscopy (CD) measurement of refolded CPY and RNase B indicated that the structure of deglycosylated proteins had recovered their native status. This method offers the possibility of efficiently releasing N-linked glycans from glycoproteins and obtaining the native target proteins.