Targeted mutations in Val101 and Arg27 interferon beta protein increase its transcriptional and translational activities

Kay, Maryam; Hasan‐Abad, Amin Moradi; Hojati, Zohreh; Korbekandi, Hasan

doi:10.1016/j.cyto.2015.11.019

Cited by 6 publications

(10 citation statements)

References 20 publications

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“…On the other hand, the therapeutic effects of these mutations are also remarkable in such a way to not only improve the half-life of the mHuIFN-β in the body, but also decrease its immunogenicity. 6,19,20 The mutation (R27T) produces a new glycosylation signal on rHuIFN-β and glycosylation of the Asn25 amino acid. The glycosylation of the Asn25 in the binding of rHuIFNβ-1a to the receptor not only does not inhibit, but also by creating a new hydrogen bond relationship between Asn25-linked glycan and Thr44, Asn51 of receptor IFNAR2, may lead to creating a more stable compound between the mHuIFN-β-27/IFNAR2.…”

Section: Discussionmentioning

confidence: 99%

“…6,19 The V101F mutation also increases the expression of rHuIFNβ up to several times. 6,20 Because R27 and V101 are in the region that are involved in the binding of HuIFN-β to their receptors, changes in these regions may be affected on interferon binding to their receptors. Despite the remarkable effect of these mutations to increase the solubility, half-life, activity and increasing the expression, so the effect of these mutations should be investigated for interferon binding to the receptor.…”

Section: Introductionmentioning

confidence: 98%

“…4 Today, a wide range of functions is evident in diverse biological fields such as defense against microbial infections, antiviral, anticancer, autoimmune, inflammatory, immunization, proliferation, differentiation and survival of the cell. 4-6 These small messenger molecules (kDa 30>) are secreted by various cells of the vertebrates in response to various stimuli that play an important role in maintaining health. 1,5,7 Interferons were the first cytokines that were used for therapeutic purposes as natural forms or recombinant forms in humans.…”

Section: Introductionmentioning

confidence: 99%

“…Optimizing the culture medium, optimizing growth conditions and making changes in different areas of interferon mRNA, were causes to increase not only the expression but also the stability of HuIFN-β. 2,6 The creation of the R27T mutation causes the formation of a new N-glycosylation site on the twenty-fifth amino acid (Asn25), which significantly increases the solubility and the half-life of rHuIFN-β. 6,19 The V101F mutation also increases the expression of rHuIFNβ up to several times.…”

Section: Introductionmentioning

confidence: 99%

“…2,6 The creation of the R27T mutation causes the formation of a new N-glycosylation site on the twenty-fifth amino acid (Asn25), which significantly increases the solubility and the half-life of rHuIFN-β. 6,19 The V101F mutation also increases the expression of rHuIFNβ up to several times. 6,20 Because R27 and V101 are in the region that are involved in the binding of HuIFN-β to their receptors, changes in these regions may be affected on interferon binding to their receptors.…”

Section: Introductionmentioning

confidence: 99%

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The Comparison Between the Mutated HuIFN-β 27-101 and the WildType Interferon β: the Comprehensive In Silico Study to Evaluate theEffect of Mutations on IFN-β

Balkhi

Hojati

2019

Adv Pharm Bull

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Purpose: Interferon beta (IFN-β) is used to combat multiple sclerosis (MS) disease. Creating R27T and V101F mutations (mHuIFN-β-27 and mHuIFN-β-101) is one of the tasks performed to improve human interferon beta (HuIFN-β) half-life, function and expression. In this work, the impact of R27T and V101F mutations in recombinant IFN-β on its binding to interferon receptors were studied by molecular docking. Methods: This work was performed through in silico study. The simulation of mutation was performed using the online Rosetta Backrub software and checked using server verify3D. Comparison of access to the solvent of the amino acids in the structures created was performed using the asaview online server. Also, the effect of mutations on the fold of the protein was reviewed by the online HOPE server. The molecular docking was performed between HuIFN-β and the external region of IFNAR receptor using the online ClusPro2 protein-protein docking server. Results: The comparison of the values of the negative binding energy (ΔGbind) obtained from protein-protein molecular docking between IFNAR receptor and HuIFN-β, mHuIFN-β-27, mHuIFN-β-101 and mHuIFN-β-27-101 ligands did not show a significant difference, and these differences do not see any meaningful relationship between them (P > 0.9999). Conclusion: Regarding these results, it can be concluded that these mutations do not have a negative effect on the composition of the complex rHuIFN-β/IFNAR. So, they do not interfere with the binding of the IFN-β to the receptor. It is concluded that the quality of the rHuIFN-β is improved by introducing these two mutations.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The Comparison Between the Mutated HuIFN-β 27-101 and the WildType Interferon β: the Comprehensive In Silico Study to Evaluate theEffect of Mutations on IFN-β

Balkhi

Hojati

2019

Adv Pharm Bull

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Plant‐Based Anticancer Compounds With a Focus on Breast Cancer

Hasan‐Abad,

Atapour,

Sobhani‐Nasab

et al. 2024

Cancer Reports

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Breast cancer is a common form of cancer among women characterized by the growth of malignant cells in the breast tissue. The most common treatments for this condition include chemotherapy, surgical intervention, radiation therapy, hormone therapy, and biological therapy. The primary issues associated with chemotherapy and radiation therapy are their adverse events and significant financial burden among patients in underdeveloped countries. This highlights the need to explore and develop superior therapeutic options that are less detrimental and more economically efficient. Plants provide an abundant supply of innovative compounds and present a promising new avenue for investigating cancer. Plants and their derivations are undergoing a revolution due to their reduced toxicity, expediency, cost‐effectiveness, safety, and simplicity in comparison to conventional treatment methods. Natural products are considered promising candidates for the development of anticancer drugs, due perhaps to the diverse pleiotropic effects on target events. The effects of plant‐derived products are limited to cancer cells while leaving healthy cells unaffected. Identification of compounds with strong anticancer properties and development of plant‐based medications for cancer treatment might be crucial steps in breast cancer therapy. Although bioactive compounds have potent anticancer properties, they also have drawbacks that need to be resolved before their application in clinical trials and improved for the approved drugs. This study aims to give comprehensive information on known anticancer compounds, including their sources and molecular mechanisms of actions, along with opportunities and challenges in plant‐based anticancer therapies.

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In vitro and computational studies on the effects of ARE deletion and targeted mutations on the expression of interferon beta-1a in HEK293T cells

Norouzi

Hojati

Dehbashi

2018

Appl Microbiol Biotechnol

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Interferon beta (IFNβ) is transiently expressed in response to viral infections and widely used to treat relapsing-remitting multiple sclerosis (MS). We introduced mutations in the IFNβ gene (in the 27th and 101st codons and in the Kozak sequence, and also deletion of 3' and 5' unstable, untranslated region, UTR) with the aim of increasing the expression of IFNβ. Computational analyses of mutant and wild-type RNAs and proteins of IFNβ by RNAfold, ASAView, HOPE and Ramachandran plot, and iStable web servers showed that the mutations could decrease RNA stability, protein solvent accessibility, and protein stability but could not change correct folding. Two recombinant IFNβ101 and IFNβ101+27 constructs were designed by site-directed mutagenesis. The wild-type IFNβ gene also was used as a control. In vitro experiments by quantitative real-time PCR, dot blot, SDS-PAGE, and Western blot assays showed an increased expression level of recombinant IFNβs. 79.9-fold, 99.7-fold, and 190-fold elevations in the mRNA expression of IFNβw, IFNβ101, and IFNβ101+27 were seen, respectively, in comparison with the endogenous IFNβ mRNA in untransfected HEK293T cells. The IFNβ mRNA expression was increased 2.38-fold and 1.25-fold for 101+27 and 101 mutated forms, respectively, in comparison with the IFNβ wild-type construct. An elevation in IFNβ protein production was also clearly detected in the transfected HEK293T cell containing recombinant IFNβ101 and IFNβ101+27 constructs. Finally, these directed mutations in the IFNβ gene successfully elevated protein and mRNA production but in silico analyses of mutant mRNAs showed decreased mRNA stability, unlike previous studies, in comparison with the wild-type mRNA.

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Targeted mutations in Val101 and Arg27 interferon beta protein increase its transcriptional and translational activities

Cited by 6 publications

References 20 publications

The Comparison Between the Mutated HuIFN-β 27-101 and the WildType Interferon β: the Comprehensive In Silico Study to Evaluate theEffect of Mutations on IFN-β

The Comparison Between the Mutated HuIFN-β 27-101 and the WildType Interferon β: the Comprehensive In Silico Study to Evaluate theEffect of Mutations on IFN-β

Plant‐Based Anticancer Compounds With a Focus on Breast Cancer

In vitro and computational studies on the effects of ARE deletion and targeted mutations on the expression of interferon beta-1a in HEK293T cells

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