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Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum

Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain of P. ox...

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Autores principales: Singh, Mangaljeet, Singh, Harpreet, Kaur, Kirandeep, Shubhankar, Shubhankar, Singh, Supreet, Kaur, Amarjeet, Singh, Prabhjeet
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10575979/
https://www.ncbi.nlm.nih.gov/pubmed/37833355
http://dx.doi.org/10.1038/s41598-023-44606-5
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author Singh, Mangaljeet
Singh, Harpreet
Kaur, Kirandeep
Shubhankar, Shubhankar
Singh, Supreet
Kaur, Amarjeet
Singh, Prabhjeet
author_facet Singh, Mangaljeet
Singh, Harpreet
Kaur, Kirandeep
Shubhankar, Shubhankar
Singh, Supreet
Kaur, Amarjeet
Singh, Prabhjeet
author_sort Singh, Mangaljeet
collection PubMed
description Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain of P. oxalicum was associated with enhanced expression of a cyclophilin gene, PoxCYP18. Cloning and characterization of PoxCYP18 revealed that its cDNA consists of 522 bp encoding a protein of 173 amino acid residues, with predicted molecular mass and pI values of 18.91 kDa and 8.87, respectively. The recombinant PoxCYP18 can catalyze cis–trans isomerization of peptidyl-prolyl bond with a catalytic efficiency of 1.46 × 10(7) M(−1) s(−1) and is inhibited specifically only by cyclosporin A, with an inhibition constant of 5.04 ± 1.13 nM. PoxCYP18 consists of two cysteine residues at positions − 45 and − 170, and loses its activity under oxidizing conditions. Substitution of these residues alone or together by site-directed mutagenesis revealed that the PPIase activity of PoxCYP18 is regulated through a redox mechanism involving the formation of disulfide linkages. Heterologous expression of PoxCYP18 conferred enhanced tolerance to salt stress in transgenic E. coli cells, implying that this protein imparts protection to cellular processes against salt-induced damage.
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spelling pubmed-105759792023-10-15 Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum Singh, Mangaljeet Singh, Harpreet Kaur, Kirandeep Shubhankar, Shubhankar Singh, Supreet Kaur, Amarjeet Singh, Prabhjeet Sci Rep Article Penicillium species are an industrially important group of fungi. Cyclophilins are ubiquitous proteins and several members of this family exhibit peptidyl-prolyl cis–trans isomerase (PPIase) activity. We had earlier demonstrated that the salt-induced PPIase activity in a halotolerant strain of P. oxalicum was associated with enhanced expression of a cyclophilin gene, PoxCYP18. Cloning and characterization of PoxCYP18 revealed that its cDNA consists of 522 bp encoding a protein of 173 amino acid residues, with predicted molecular mass and pI values of 18.91 kDa and 8.87, respectively. The recombinant PoxCYP18 can catalyze cis–trans isomerization of peptidyl-prolyl bond with a catalytic efficiency of 1.46 × 10(7) M(−1) s(−1) and is inhibited specifically only by cyclosporin A, with an inhibition constant of 5.04 ± 1.13 nM. PoxCYP18 consists of two cysteine residues at positions − 45 and − 170, and loses its activity under oxidizing conditions. Substitution of these residues alone or together by site-directed mutagenesis revealed that the PPIase activity of PoxCYP18 is regulated through a redox mechanism involving the formation of disulfide linkages. Heterologous expression of PoxCYP18 conferred enhanced tolerance to salt stress in transgenic E. coli cells, implying that this protein imparts protection to cellular processes against salt-induced damage. Nature Publishing Group UK 2023-10-13 /pmc/articles/PMC10575979/ /pubmed/37833355 http://dx.doi.org/10.1038/s41598-023-44606-5 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Singh, Mangaljeet
Singh, Harpreet
Kaur, Kirandeep
Shubhankar, Shubhankar
Singh, Supreet
Kaur, Amarjeet
Singh, Prabhjeet
Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
title Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
title_full Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
title_fullStr Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
title_full_unstemmed Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
title_short Characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of Penicillium oxalicum
title_sort characterization and regulation of salt upregulated cyclophilin from a halotolerant strain of penicillium oxalicum
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10575979/
https://www.ncbi.nlm.nih.gov/pubmed/37833355
http://dx.doi.org/10.1038/s41598-023-44606-5
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