Yeast Heterologous Expression Systems for the Study of Plant Membrane Proteins

Abstract

Researchers are often interested in proteins that are present in cells in small ratios compared to the total amount of proteins. These proteins include transcription factors, hormones and specific membrane proteins. However, sufficient amounts of well-purified protein preparations are required for functional and structural studies of these proteins, including the creation of artificial proteoliposomes and the growth of protein 2D and 3D crystals. This aim can be achieved by the expression of the target protein in a heterologous system. This review describes the applications of yeast heterologous expression systems in studies of plant membrane proteins. An initial brief description introduces the widely used heterologous expression systems of the baker’s yeast Saccharomyces cerevisiae and the methylotrophic yeast Pichia pastoris. S. cerevisiae is further considered a convenient model system for functional studies of heterologously expressed proteins, while P. pastoris has the advantage of using these yeast cells as factories for producing large quantities of proteins of interest. The application of both expression systems is described for functional and structural studies of membrane proteins from plants, namely, K+- and Na+-transporters, various ATPases and anion transporters, and other transport proteins.

Evaluation of heterologous expression in Pichia pastoris of Pine Weevil TRPA1 by GFP and flow cytometry

“…4). Other membrane proteins have been reported to localize both to the plasma membrane and in internal structures [22][23][24][25] or specifically to accumulate in stacked membranes and perinuclear membranes [26]. It has been proposed that a preferentially internal localization would correlate with protein size and/or the number of transmembrane helices [22].…”

Cloning and Overexpressing Membrane Proteins Using Pichia pastoris (Komagataella phaffii)

“…Usually there are two scenarios: no information is available in the literature, or similar proteins have already been successfully produced in this system and the published data can serve as a useful starting point. In the latter case, it is advisable to consult the few authoritative surveys that have been conducted on various IMPs expressed in P. pastoris (Alkhalfioui et al, 2011;Bill & Hedfalk, 2021;Byrne, 2015;Hausjell et al, 2020;Popova et al, 2023). When no published information is available, a combination of vectors that comprise the following sequence elements must be considered.…”