LAUSR

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.