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Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders
Generally, in vivo function and structural changes are studied by probing proteins in a dilute solution under in vitro conditions, which is believed to be mimicking proteins in intracellular milieu. Earlier, thermal-induced denaturation of myoglobin, in the milieu of crowder molecule showed destabil...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7175338/ https://www.ncbi.nlm.nih.gov/pubmed/32210191 http://dx.doi.org/10.3390/biom10030490 |
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author | Nasreen, Khalida Parray, Zahoor Ahmad Ahamad, Shahzaib Ahmad, Faizan Ahmed, Anwar Freeh Alamery, Salman Hussain, Tajamul Hassan, Md. Imtaiyaz Islam, Asimul |
author_facet | Nasreen, Khalida Parray, Zahoor Ahmad Ahamad, Shahzaib Ahmad, Faizan Ahmed, Anwar Freeh Alamery, Salman Hussain, Tajamul Hassan, Md. Imtaiyaz Islam, Asimul |
author_sort | Nasreen, Khalida |
collection | PubMed |
description | Generally, in vivo function and structural changes are studied by probing proteins in a dilute solution under in vitro conditions, which is believed to be mimicking proteins in intracellular milieu. Earlier, thermal-induced denaturation of myoglobin, in the milieu of crowder molecule showed destabilization of the metal protein. Destabilization of protein by thermal-induced denaturation involves a large extrapolation, so, the reliability is questionable. This led us to measure the effects of macromolecular crowding on its stability by chemical-induced denaturation of the protein using probes like circular dichroism and absorption spectroscopy in the presence of dextran 70 and ficoll 70 at various pHs (acidic: 6.0, almost neutral: 7.0 and basic: 8.0). Observations showed that the degree of destabilization of myoglobin was greater due to ficoll 70 as compared to that of dextran 70 so it can be understood that the nature of the crowder or the shape of the crowder has an important role towards the stability of proteins. Additionally, the degree of destabilization was observed as pH dependent, however the pH dependence is different for different crowders. Furthermore, isothermal titration calorimetry and molecular docking studies confirmed that both the crowders (ficoll and dextran) bind to heme moiety of myoglobin and a single binding site was observed for each. |
format | Online Article Text |
id | pubmed-7175338 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-71753382020-04-28 Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders Nasreen, Khalida Parray, Zahoor Ahmad Ahamad, Shahzaib Ahmad, Faizan Ahmed, Anwar Freeh Alamery, Salman Hussain, Tajamul Hassan, Md. Imtaiyaz Islam, Asimul Biomolecules Article Generally, in vivo function and structural changes are studied by probing proteins in a dilute solution under in vitro conditions, which is believed to be mimicking proteins in intracellular milieu. Earlier, thermal-induced denaturation of myoglobin, in the milieu of crowder molecule showed destabilization of the metal protein. Destabilization of protein by thermal-induced denaturation involves a large extrapolation, so, the reliability is questionable. This led us to measure the effects of macromolecular crowding on its stability by chemical-induced denaturation of the protein using probes like circular dichroism and absorption spectroscopy in the presence of dextran 70 and ficoll 70 at various pHs (acidic: 6.0, almost neutral: 7.0 and basic: 8.0). Observations showed that the degree of destabilization of myoglobin was greater due to ficoll 70 as compared to that of dextran 70 so it can be understood that the nature of the crowder or the shape of the crowder has an important role towards the stability of proteins. Additionally, the degree of destabilization was observed as pH dependent, however the pH dependence is different for different crowders. Furthermore, isothermal titration calorimetry and molecular docking studies confirmed that both the crowders (ficoll and dextran) bind to heme moiety of myoglobin and a single binding site was observed for each. MDPI 2020-03-23 /pmc/articles/PMC7175338/ /pubmed/32210191 http://dx.doi.org/10.3390/biom10030490 Text en © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Nasreen, Khalida Parray, Zahoor Ahmad Ahamad, Shahzaib Ahmad, Faizan Ahmed, Anwar Freeh Alamery, Salman Hussain, Tajamul Hassan, Md. Imtaiyaz Islam, Asimul Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders |
title | Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders |
title_full | Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders |
title_fullStr | Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders |
title_full_unstemmed | Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders |
title_short | Interactions Under Crowding Milieu: Chemical-Induced Denaturation of Myoglobin is Determined by the Extent of Heme Dissociation on Interaction with Crowders |
title_sort | interactions under crowding milieu: chemical-induced denaturation of myoglobin is determined by the extent of heme dissociation on interaction with crowders |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7175338/ https://www.ncbi.nlm.nih.gov/pubmed/32210191 http://dx.doi.org/10.3390/biom10030490 |
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