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Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities

BACKGROUND: Ocriplasmin has been developed for the induction of posterior vitreous detachment in patients with vitreomacular adhesion. At physiological pH, ocriplasmin is susceptible to autolytic and proteolytic degradation, limiting its activity duration. These undesirable properties of ocriplasmin...

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Autores principales: Baghban, Roghayyeh, Farajnia, Safar, Ghasemi, Younes, Mortazavi, Mojtaba, Ghasemali, Samaneh, Zakariazadeh, Mostafa, Zarghami, Nosratollah, Samadi, Nasser
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Shiraz University of Medical Sciences 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8611222/
https://www.ncbi.nlm.nih.gov/pubmed/34840386
http://dx.doi.org/10.30476/ijms.2020.86984.1705
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author Baghban, Roghayyeh
Farajnia, Safar
Ghasemi, Younes
Mortazavi, Mojtaba
Ghasemali, Samaneh
Zakariazadeh, Mostafa
Zarghami, Nosratollah
Samadi, Nasser
author_facet Baghban, Roghayyeh
Farajnia, Safar
Ghasemi, Younes
Mortazavi, Mojtaba
Ghasemali, Samaneh
Zakariazadeh, Mostafa
Zarghami, Nosratollah
Samadi, Nasser
author_sort Baghban, Roghayyeh
collection PubMed
description BACKGROUND: Ocriplasmin has been developed for the induction of posterior vitreous detachment in patients with vitreomacular adhesion. At physiological pH, ocriplasmin is susceptible to autolytic and proteolytic degradation, limiting its activity duration. These undesirable properties of ocriplasmin can be reduced by site-directed mutagenesis, so that its enzymatic activities can be augmented. This study aimed to design ocriplasmin variants with improved biological/physicochemical characteristics via bioinformatics tools. METHODS: This study was performed in Tabriz University of Medical Sciences, Tabriz, Iran, 2019. Through site-directed mutagenesis, three ocriplasmin variants were designed. Structural analysis was performed on the wild-type variant and the mutant variants using the Protein Interactions Calculator (PIC) server. The interactions between the S-2403 substrate and the ocriplasmin variants were studied by molecular docking simulations, and binding capability was evaluated by the calculation of free binding energy. The conformational features of protein-substrate complex systems for all the variants were evaluated using molecular dynamic simulations at 100 nanoseconds. RESULTS: The structural analysis of ocriplasmin revealed that the substitution of threonine for alanine 59 significantly reduced proteolytic activity, while the substitution of glutamic acid for lysine 156 influenced autolytic function. The molecular docking simulation results indicated the appropriate binding of the substrate to the ocriplasmin variants with high-to-low affinities. The binding affinity of the wild-type variant for the substrate was higher than that between the mutant variants and the substrate. Simulation analyses, consisting of the root-mean-square deviation, the root-mean-square fluctuation, and the center-of-mass average distance showed a higher affinity of the substrate for the wild type than for the mutant variants. CONCLUSION: The mutational analysis of ocriplasmin revealed that A59T and K156E mutagenesis could be used for the development of a new variant with higher therapeutic efficacy.
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spelling pubmed-86112222021-11-26 Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities Baghban, Roghayyeh Farajnia, Safar Ghasemi, Younes Mortazavi, Mojtaba Ghasemali, Samaneh Zakariazadeh, Mostafa Zarghami, Nosratollah Samadi, Nasser Iran J Med Sci Original Article BACKGROUND: Ocriplasmin has been developed for the induction of posterior vitreous detachment in patients with vitreomacular adhesion. At physiological pH, ocriplasmin is susceptible to autolytic and proteolytic degradation, limiting its activity duration. These undesirable properties of ocriplasmin can be reduced by site-directed mutagenesis, so that its enzymatic activities can be augmented. This study aimed to design ocriplasmin variants with improved biological/physicochemical characteristics via bioinformatics tools. METHODS: This study was performed in Tabriz University of Medical Sciences, Tabriz, Iran, 2019. Through site-directed mutagenesis, three ocriplasmin variants were designed. Structural analysis was performed on the wild-type variant and the mutant variants using the Protein Interactions Calculator (PIC) server. The interactions between the S-2403 substrate and the ocriplasmin variants were studied by molecular docking simulations, and binding capability was evaluated by the calculation of free binding energy. The conformational features of protein-substrate complex systems for all the variants were evaluated using molecular dynamic simulations at 100 nanoseconds. RESULTS: The structural analysis of ocriplasmin revealed that the substitution of threonine for alanine 59 significantly reduced proteolytic activity, while the substitution of glutamic acid for lysine 156 influenced autolytic function. The molecular docking simulation results indicated the appropriate binding of the substrate to the ocriplasmin variants with high-to-low affinities. The binding affinity of the wild-type variant for the substrate was higher than that between the mutant variants and the substrate. Simulation analyses, consisting of the root-mean-square deviation, the root-mean-square fluctuation, and the center-of-mass average distance showed a higher affinity of the substrate for the wild type than for the mutant variants. CONCLUSION: The mutational analysis of ocriplasmin revealed that A59T and K156E mutagenesis could be used for the development of a new variant with higher therapeutic efficacy. Shiraz University of Medical Sciences 2021-11 /pmc/articles/PMC8611222/ /pubmed/34840386 http://dx.doi.org/10.30476/ijms.2020.86984.1705 Text en Copyright: © Iranian Journal of Medical Sciences https://creativecommons.org/licenses/by-nd/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 4.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Baghban, Roghayyeh
Farajnia, Safar
Ghasemi, Younes
Mortazavi, Mojtaba
Ghasemali, Samaneh
Zakariazadeh, Mostafa
Zarghami, Nosratollah
Samadi, Nasser
Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities
title Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities
title_full Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities
title_fullStr Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities
title_full_unstemmed Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities
title_short Engineering of Ocriplasmin Variants by Bioinformatics Methods for the Reduction of Proteolytic and Autolytic Activities
title_sort engineering of ocriplasmin variants by bioinformatics methods for the reduction of proteolytic and autolytic activities
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8611222/
https://www.ncbi.nlm.nih.gov/pubmed/34840386
http://dx.doi.org/10.30476/ijms.2020.86984.1705
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