Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/100244
Title: Expression and purification of coronavirus envelope proteins using a modified β-barrel construct
Authors: Pervushin, Konstantin
Torres, Jaume
Parthasarathy, Krupakar
Lu, Huang
Surya, Wahyu
Vararattanavech, Ardcharaporn
Keywords: DRNTU::Science::Biological sciences
Issue Date: 2012
Source: Parthasarathy, K., Lu, H., Surya, W., Vararattanavech, A., Pervushin, K., & Torres, J. (2012). Expression and purification of coronavirus envelope proteins using a modified β-barrel construct. Protein Expression and Purification, 85(1), 133–141.
Series/Report no.: Protein expression and purification
Abstract: Coronavirus envelope (E) proteins are short (∼100 residues) polypeptides that contain at least one transmembrane (TM) domain and a cluster of 2–3 juxtamembrane cysteines. These proteins are involved in viral morphogenesis and tropism, and their absence leads in some cases to aberrant virions, or to viral attenuation. In common to other viroporins, coronavirus envelope proteins increase membrane permeability to ions. Although an NMR-based model for the TM domain of the E protein in the severe acute respiratory syndrome virus (SARS-CoV E) has been reported, structural data and biophysical studies of full length E proteins are not available because efficient expression and purification methods for these proteins are lacking. Herein we have used a novel fusion protein consisting of a modified β-barrel to purify both wild type and cysteine-less mutants of two representatives of coronavirus E proteins: the shortest (76 residues), from SARS-CoV E, and one of the longest (109 residues), from the infectious bronchitis virus (IBV E). The fusion construct was subsequently cleaved with cyanogen bromide and all polypeptides were obtained with high purity. This is an approach that can be used in other difficult hydrophobic peptides.
URI: https://hdl.handle.net/10356/100244
http://hdl.handle.net/10220/10981
ISSN: 1046-5928
Rights: © 2012 Elsevier Inc.
Fulltext Permission: none
Fulltext Availability: No Fulltext
Appears in Collections:SBS Journal Articles

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