CAS Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China; Shandong Engineering Laboratory of Single Cell Oil, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 266101, China; Shandong Energy Institute, Qingdao, Shandong 266101, China; Qingdao New Energy Shandong Laboratory, Qingdao, Shandong 266101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Int J Biol Macromol. 2023 Dec 31;253(Pt 1):126649. doi: 10.1016/j.ijbiomac.2023.126649. Epub 2023 Sep 2.
There is an increasing interest in using S-glycosylation as a replacement for the more commonly occurring O-glycosylation, aiming to enhance the resistance of glycans against chemical hydrolysis and enzymatic degradation. However, previous studies have demonstrated that these two types of glycosylation exert distinct effects on protein properties and functions. In order to elucidate the structural basis behind the observed differences, we conducted a systematic and comparative analysis of 6 differently glycosylated forms of a model glycoprotein, CBM, using NMR spectroscopy and molecular dynamic simulations. Our findings revealed that the different stabilizing effects of S- and O-glycosylation could be attributed to altered hydrogen-bonding capability between the glycan and the polypeptide chain, and their diverse impacts on binding affinity could be elucidated by examining the interactions and motion dynamics of glycans in substrate-bound states. Overall, this study underscores the pivotal role of the glycosidic linkage in shaping the function of glycosylation and advises caution when switching glycosylation types in protein glycoengineering.
人们越来越感兴趣地使用 S-糖基化来替代更常见的 O-糖基化，旨在提高聚糖对化学水解和酶降解的抗性。然而，以前的研究表明，这两种类型的糖基化对蛋白质的性质和功能有不同的影响。为了阐明观察到的差异的结构基础，我们使用 NMR 光谱和分子动力学模拟对一种模型糖蛋白 CBM 的 6 种不同糖基化形式进行了系统和比较分析。我们的研究结果表明，S-和 O-糖基化的不同稳定作用可以归因于聚糖和多肽链之间氢键结合能力的改变，并且可以通过研究底物结合状态下糖基的相互作用和运动动力学来阐明它们对结合亲和力的不同影响。总的来说，这项研究强调了糖苷键在塑造糖基化功能中的关键作用，并建议在糖基工程中切换糖基化类型时要谨慎。
