Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78

Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78 Publisher Pubmed

Aghamollaei H¹ ; Mousavi Gargari SL² ; Ghanei M^{3, 7} ; Rasaee MJ⁴ ; Amani J⁵ ; Bakherad H⁶ ; Farnoosh G¹

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1. Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
2. Department of Biology, Faculty of Basic Science, Shahed University, Tehran, Iran
3. Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
4. Medical Biotechnology Department, Faculty of Medical sciences, Tarbiat Modares University, Tehran, Iran
5. Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
6. Department of Pharmaceutical Biotechnology, Faculty of pharmacy, Tehran University of Medical Sciences, Tehran, Iran
7. Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Molasadra Street, Vanak Square, Tehran, Iran

Source: Biotechnology and Applied Biochemistry Published:2017

Abstract

Glucose-regulated protein 78 (GRP78) is a typical endoplasmic reticulum luminal chaperone having a main role in the activation of the unfolded protein response. Because of hypoxia and nutrient deprivation in the tumor microenvironment, expression of GRP78 in these cells becomes higher than the native cells, which makes it a suitable candidate for cancer targeting. Suppression of survival signals by antibody production against C-terminal domain of GR78 (CGRP) can induce apoptosis of cancer cells. The aim of this study was in silico analysis, recombinant production, and characterization of CGRP in Escherichia coli. Structural prediction of CGRP by bioinformatics tools was done and the construct containing optimized sequence was transferred to E. coli T7 shuffle. Expression was induced by isopropyl-β-d-thiogalactoside, and recombinant protein was purified by Ni-NTA agarose resin. The content of secondary structures was obtained by circular dichroism (CD) spectrum. CGRP immunogenicity was evaluated from the immunized mouse sera. SDS-PAGE analysis showed CGRP expression in E. coli. CD spectrum also confirmed prediction of structures by bioinformatics tools. The enzyme-linked immunosorbent assay using sera from immunized mice revealed CGRP as a good immunogen. The results obtained in this study showed that the structure of truncated CGRP is very similar to its structure in the whole protein context. This protein can be used in cancer researches. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

Style	Citing Format
MLA	Aghamollaei H, et al.. "Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78." Biotechnology and Applied Biochemistry, vol. 64, no. 1, 2017, pp. 117-125.
APA	Aghamollaei H, Mousavi Gargari SL, Ghanei M, Rasaee MJ, Amani J, Bakherad H, Farnoosh G (2017). Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78. Biotechnology and Applied Biochemistry, 64(1), 117-125.
Chicago	Aghamollaei H, Mousavi Gargari SL, Ghanei M, Rasaee MJ, Amani J, Bakherad H, Farnoosh G. "Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78." Biotechnology and Applied Biochemistry 64, no. 1 (2017): 117-125.
Harvard	Aghamollaei H et al. (2017) 'Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78', Biotechnology and Applied Biochemistry, 64(1), pp. 117-125.
Vancouver	Aghamollaei H, Mousavi Gargari SL, Ghanei M, Rasaee MJ, Amani J, Bakherad H, et al.. Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78. Biotechnology and Applied Biochemistry. 2017;64(1):117-125.
BibTex	@article{ author = {Aghamollaei H and Mousavi Gargari SL and Ghanei M and Rasaee MJ and Amani J and Bakherad H and Farnoosh G}, title = {Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78}, journal = {Biotechnology and Applied Biochemistry}, volume = {64}, number = {1}, pages = {117-125}, year = {2017} }
RIS	TY - JOUR AU - Aghamollaei H AU - Mousavi Gargari SL AU - Ghanei M AU - Rasaee MJ AU - Amani J AU - Bakherad H AU - Farnoosh G TI - Structure Prediction, Expression, and Antigenicity of C-Terminal of Grp78 JO - Biotechnology and Applied Biochemistry VL - 64 IS - 1 SP - 117 EP - 125 PY - 2017 ER -

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