Identifying Stable Fragments of Arabidopsis thaliana Cellulose Synthase Subunit 3 by Yeast Display. Issue 4 (25th September 2018)

• Record Type: Journal Article
• Authors: Raeeszadeh‐Sarmazdeh, Maryam; Patel, Nikhil; Cruise, Sarah; Owen, Leila; O'Neill, Hugh; Boder, Eric T.
• Is Part Of: Biotechnology journal. Volume 14:Issue 4(2019)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-09-25
• Subjects: cellulose synthase -- domain identification -- library of random protein fragments -- protein stability -- yeast surface display; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201800353
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract : 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. Abstract : Yeast surface display (YSD) is used to determine full‐length expression of Arabidopsis thaliana cellulose synthase 3 (AtCesA3) fragments. A library of random AtCesA3 fragments displayed on the yeast surface is screened based on full‐length expression level using fluorescent‐activated cell sorting (FACS). This technique provides a fast and simple method for screening protein domains and fragments of complex proteins to identify potential stable protein fragments.