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Identifying Stable Fragments of Arabidopsis thaliana Cellulose Synthase Subunit 3 by Yeast Display
Author(s) -
RaeeszadehSarmazdeh Maryam,
Patel Nikhil,
Cruise Sarah,
Owen Leila,
O'Neill Hugh,
Boder Eric T.
Publication year - 2019
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201800353
Subject(s) - yeast , arabidopsis thaliana , protein subunit , atp synthase , arabidopsis , cellulose , chemistry , biochemistry , biology , enzyme , gene , mutant
Determining structures of large, complex proteins remains challenging, especially for transmembrane proteins, as the protein size increases. Arabidopsis thaliana cellulose synthesis complex is a large, multimeric complex located in the plant cell membrane that synthesizes cellulose microfibrils in the plant cell wall. Despite the biological and economic importance of cellulose and therefore cellulose synthesis, many aspects of the cellulase synthase complex (CSC) structure and function are still unknown. Here, yeast surface display (YSD) is used to determine the full‐length expression of A. thaliana cellulose synthase 3 (AtCesA3) fragments. The level of stably‐folded AtCesA3 fragments displayed on the yeast surface are determined using flow cytometric analysis of differential surface expression of epitopes flanking the AtCesA3 fragment. This technique provides a fast and simple method for examining folding and expression of protein domains and fragments of complex proteins.