Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Structure prediction, expression, and antigenicity of c-terminal of GRP78

(2017) Structure prediction, expression, and antigenicity of c-terminal of GRP78. Biotechnology and Applied Biochemistry. pp. 117-125. ISSN 0885-4513

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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--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.

Item Type: Article
Keywords: bioinformatics cancer GRP78 prediction recombinant glucose-regulated protein endoplasmic-reticulum chaperone secondary structure prediction stress signaling pathway cell-surface grp78 rheumatoid-arthritis escherichia-coli prostate-cancer t-cells tumor Biochemistry & Molecular Biology Biotechnology & Applied Microbiology
Page Range: pp. 117-125
Journal or Publication Title: Biotechnology and Applied Biochemistry
Journal Index: ISI
Volume: 64
Number: 1
Identification Number:
ISSN: 0885-4513
Depositing User: مهندس مهدی شریفی

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