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Structural and Functional Differences between Homologous Bacterial Ribonucleases
Small cationic guanyl-preferring ribonucleases (RNases) produced by the Bacillus species share a similar protein tertiary structure with a high degree of amino acid sequence conservation. However, they form dimers that differ in conformation and stability. Here, we have addressed the issues (1) whet...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8837141/ https://www.ncbi.nlm.nih.gov/pubmed/35163789 http://dx.doi.org/10.3390/ijms23031867 |
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author | Ulyanova, Vera Nadyrova, Alsu Dudkina, Elena Kuznetsova, Aleksandra Ahmetgalieva, Albina Faizullin, Dzhigangir Surchenko, Yulia Novopashina, Darya Zuev, Yuriy Kuznetsov, Nikita Ilinskaya, Olga |
author_facet | Ulyanova, Vera Nadyrova, Alsu Dudkina, Elena Kuznetsova, Aleksandra Ahmetgalieva, Albina Faizullin, Dzhigangir Surchenko, Yulia Novopashina, Darya Zuev, Yuriy Kuznetsov, Nikita Ilinskaya, Olga |
author_sort | Ulyanova, Vera |
collection | PubMed |
description | Small cationic guanyl-preferring ribonucleases (RNases) produced by the Bacillus species share a similar protein tertiary structure with a high degree of amino acid sequence conservation. However, they form dimers that differ in conformation and stability. Here, we have addressed the issues (1) whether the homologous RNases also have distinctions in catalytic activity towards different RNA substrates and interactions with the inhibitor protein barstar, and (2) whether these differences correlate with structural features of the proteins. Circular dichroism and dynamic light scattering assays revealed distinctions in the structures of homologous RNases. The activity levels of the RNases towards natural RNA substrates, as measured spectrometrically by acid-soluble hydrolysis products, were similar and decreased in the row high-polymeric RNA >>> transport RNA > double-stranded RNA. However, stopped flow kinetic studies on model RNA substrates containing the guanosine residue in a hairpin stem or a loop showed that the cleavage rates of these enzymes were different. Moreover, homologous RNases were inhibited by the barstar with diverse efficiency. Therefore, minor changes in structure elements of homologous proteins have a potential to significantly effect molecule stability and functional activities, such as catalysis or ligand binding. |
format | Online Article Text |
id | pubmed-8837141 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-88371412022-02-12 Structural and Functional Differences between Homologous Bacterial Ribonucleases Ulyanova, Vera Nadyrova, Alsu Dudkina, Elena Kuznetsova, Aleksandra Ahmetgalieva, Albina Faizullin, Dzhigangir Surchenko, Yulia Novopashina, Darya Zuev, Yuriy Kuznetsov, Nikita Ilinskaya, Olga Int J Mol Sci Article Small cationic guanyl-preferring ribonucleases (RNases) produced by the Bacillus species share a similar protein tertiary structure with a high degree of amino acid sequence conservation. However, they form dimers that differ in conformation and stability. Here, we have addressed the issues (1) whether the homologous RNases also have distinctions in catalytic activity towards different RNA substrates and interactions with the inhibitor protein barstar, and (2) whether these differences correlate with structural features of the proteins. Circular dichroism and dynamic light scattering assays revealed distinctions in the structures of homologous RNases. The activity levels of the RNases towards natural RNA substrates, as measured spectrometrically by acid-soluble hydrolysis products, were similar and decreased in the row high-polymeric RNA >>> transport RNA > double-stranded RNA. However, stopped flow kinetic studies on model RNA substrates containing the guanosine residue in a hairpin stem or a loop showed that the cleavage rates of these enzymes were different. Moreover, homologous RNases were inhibited by the barstar with diverse efficiency. Therefore, minor changes in structure elements of homologous proteins have a potential to significantly effect molecule stability and functional activities, such as catalysis or ligand binding. MDPI 2022-02-07 /pmc/articles/PMC8837141/ /pubmed/35163789 http://dx.doi.org/10.3390/ijms23031867 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ulyanova, Vera Nadyrova, Alsu Dudkina, Elena Kuznetsova, Aleksandra Ahmetgalieva, Albina Faizullin, Dzhigangir Surchenko, Yulia Novopashina, Darya Zuev, Yuriy Kuznetsov, Nikita Ilinskaya, Olga Structural and Functional Differences between Homologous Bacterial Ribonucleases |
title | Structural and Functional Differences between Homologous Bacterial Ribonucleases |
title_full | Structural and Functional Differences between Homologous Bacterial Ribonucleases |
title_fullStr | Structural and Functional Differences between Homologous Bacterial Ribonucleases |
title_full_unstemmed | Structural and Functional Differences between Homologous Bacterial Ribonucleases |
title_short | Structural and Functional Differences between Homologous Bacterial Ribonucleases |
title_sort | structural and functional differences between homologous bacterial ribonucleases |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8837141/ https://www.ncbi.nlm.nih.gov/pubmed/35163789 http://dx.doi.org/10.3390/ijms23031867 |
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