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Title: Heterologous expression and purification of catalytic domain of CESA1 from Arabidopsis thaliana

Abstract

Here, heterologous expression of plant cellulose synthase (CESA) and its purification has remained a challenge for decades impeding detailed biophysical, biochemical and structural characterization of this key enzyme. An in-depth knowledge of structure and function of CESA proteins would enable us to better understand the hierarchical structure of the plant cell wall. Here, we report a detailed, and reproducible method of purification of catalytic domain of CESA1 from Arabidopsis thaliana that was recombinantly expressed in Escherichia coli. The method relies on a two stage purification procedure to obtain the catalytic domain in monomer and trimer forms. The biochemical and biophysical data including low resolution structures of the protein have been published. Currently the crystallization studies of this protein are underway.

Authors:
 [1];  [1]
  1. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). High Flux Isotope Reactor (HFIR); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1339410
DOE Contract Number:  
AC05-00OR22725
Resource Type:
Journal Article
Journal Name:
Bio-Protocol
Additional Journal Information:
Journal Volume: 6; Journal Issue: 20; Journal ID: ISSN 2331-8325
Publisher:
Bio-protocol LLC
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Vandavasi, Venu Gopal, and O'Neill, Hugh Michael. Heterologous expression and purification of catalytic domain of CESA1 from Arabidopsis thaliana. United States: N. p., 2016. Web. doi:10.21769/BioProtoc.1965.
Vandavasi, Venu Gopal, & O'Neill, Hugh Michael. Heterologous expression and purification of catalytic domain of CESA1 from Arabidopsis thaliana. United States. doi:10.21769/BioProtoc.1965.
Vandavasi, Venu Gopal, and O'Neill, Hugh Michael. Thu . "Heterologous expression and purification of catalytic domain of CESA1 from Arabidopsis thaliana". United States. doi:10.21769/BioProtoc.1965.
@article{osti_1339410,
title = {Heterologous expression and purification of catalytic domain of CESA1 from Arabidopsis thaliana},
author = {Vandavasi, Venu Gopal and O'Neill, Hugh Michael},
abstractNote = {Here, heterologous expression of plant cellulose synthase (CESA) and its purification has remained a challenge for decades impeding detailed biophysical, biochemical and structural characterization of this key enzyme. An in-depth knowledge of structure and function of CESA proteins would enable us to better understand the hierarchical structure of the plant cell wall. Here, we report a detailed, and reproducible method of purification of catalytic domain of CESA1 from Arabidopsis thaliana that was recombinantly expressed in Escherichia coli. The method relies on a two stage purification procedure to obtain the catalytic domain in monomer and trimer forms. The biochemical and biophysical data including low resolution structures of the protein have been published. Currently the crystallization studies of this protein are underway.},
doi = {10.21769/BioProtoc.1965},
journal = {Bio-Protocol},
issn = {2331-8325},
number = 20,
volume = 6,
place = {United States},
year = {2016},
month = {10}
}