N-linked glycosylation is one type of posttranslational modification that proteins undergo during expression. The following describes the effects of N-linked glycosylation on high-level membrane protein expression in yeast with an emphasis on Saccharomyces cerevisiae. N-linked glycosylation is highlighted here as an important consideration when preparing membrane protein gene constructs for expression in S. cerevisiae, which continues to be used as a workhorse in both research and industrial applications. Non-native N-linked glycosylation commonly occurs during the heterologous expression of mammalian proteins in many yeast species which can have important immunological consequences when used in the production of biotherapeutic proteins or peptides. Further, non-native N-linked glycosylation can lead to improper protein folding and premature degradation, which can impede high-level expression yields and hinder downstream analysis. Multiple strategies are presented in this article, which suggest different methods that can be implemented to circumvent the unwanted consequences of N-linked glycosylation during the expression process. These considerations may have long-term benefits for high-level protein production in S. cerevisiae across a broad spectrum of expression targets with special emphasis placed on G-protein coupled receptors, one of the largest families of membrane proteins.

中文翻译：

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预测的 N 端 N 连接糖基化位点可能是酿酒酵母膜蛋白表达模式的基础

N-连接的糖基化是蛋白质在表达过程中经历的一种翻译后修饰。以下描述了N-连接糖基化对酵母中高水平膜蛋白表达的影响，重点是Saccharomyces cerevisiae。在制备用于在酿酒酵母中表达的膜蛋白基因构建体时， N-连接糖基化作为一个重要的考虑因素在这里被强调，酿酒酵母继续用作研究和工业应用中的主力。非本地N连接的糖基化通常发生在哺乳动物蛋白质在许多酵母物种中的异源表达期间，当用于生产生物治疗性蛋白质或肽时，这可能具有重要的免疫学后果。此外，非天然的N-连接糖基化会导致蛋白质折叠不当和过早降解，从而阻碍高水平表达产量并阻碍下游分析。本文介绍了多种策略，这些策略提出了可以实施的不同方法，以规避表达过程中N-连接糖基化的不良后果。这些考虑可能对酿酒酵母的高水平蛋白质生产具有长期的好处涵盖广泛的表达靶点，特别强调 G 蛋白偶联受体，这是最大的膜蛋白家族之一。