Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions
The intracellular environment is overcrowded with a range of molecules (small and large), all of which influence protein conformation. As a result, understanding how proteins fold and stay functional in such crowded conditions is essential. Several in vitro experiments have looked into the effects o...
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MDPI AG
2021-05-01
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| Series: | Molecules |
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| author | Zahoor Ahmad Parray Faizan Ahmad Md. Imtaiyaz Hassan Anwar Ahmed Fahad N. Almajhdi Ajamaluddin Malik Tajamul Hussain Asimul Islam |
| author_facet | Zahoor Ahmad Parray Faizan Ahmad Md. Imtaiyaz Hassan Anwar Ahmed Fahad N. Almajhdi Ajamaluddin Malik Tajamul Hussain Asimul Islam |
| author_sort | Zahoor Ahmad Parray |
| collection | DOAJ |
| description | The intracellular environment is overcrowded with a range of molecules (small and large), all of which influence protein conformation. As a result, understanding how proteins fold and stay functional in such crowded conditions is essential. Several in vitro experiments have looked into the effects of macromolecular crowding on different proteins. However, there are hardly any reports regarding small molecular crowders used alone and in mixtures to observe their effects on the structure and stability of the proteins, which mimics of the cellular conditions. Here we investigate the effect of different mixtures of crowders, ethylene glycol (EG) and its polymer polyethylene glycol (PEG 400 Da) on the structural and thermal stability of myoglobin (Mb). Our results show that monomer (EG) has no significant effect on the structure of Mb, while the polymer disrupts its structure and decreases its stability. Conversely, the additive effect of crowders showed structural refolding of the protein to some extent. Moreover, the calorimetric binding studies of the protein showed very weak interactions with the mixture of crowders. Usually, we can assume that soft interactions induce structural perturbations while exclusion volume effects stabilize the protein structure; therefore, we hypothesize that under in vivo crowded conditions, both phenomena occur and maintain the stability and function of proteins. |
| first_indexed | 2024-03-10T11:34:50Z |
| format | Article |
| id | doaj.art-a78754bdebbc4071a24043d9a7a20196 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T11:34:50Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-a78754bdebbc4071a24043d9a7a201962023-11-21T19:00:28ZengMDPI AGMolecules1420-30492021-05-01269280710.3390/molecules26092807Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded ConditionsZahoor Ahmad Parray0Faizan Ahmad1Md. Imtaiyaz Hassan2Anwar Ahmed3Fahad N. Almajhdi4Ajamaluddin Malik5Tajamul Hussain6Asimul Islam7Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi 110025, IndiaCentre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi 110025, IndiaCentre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi 110025, IndiaCentre of Excellence in Biotechnology Research, College of Science, King Saud University, Riyadh 11451, Saudi ArabiaCentre of Excellence in Biotechnology Research, College of Science, King Saud University, Riyadh 11451, Saudi ArabiaDepartment of Biochemistry, College of Science, King Saud University, Riyadh 11451, Saudi ArabiaCentre of Excellence in Biotechnology Research, College of Science, King Saud University, Riyadh 11451, Saudi ArabiaCentre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, New Delhi 110025, IndiaThe intracellular environment is overcrowded with a range of molecules (small and large), all of which influence protein conformation. As a result, understanding how proteins fold and stay functional in such crowded conditions is essential. Several in vitro experiments have looked into the effects of macromolecular crowding on different proteins. However, there are hardly any reports regarding small molecular crowders used alone and in mixtures to observe their effects on the structure and stability of the proteins, which mimics of the cellular conditions. Here we investigate the effect of different mixtures of crowders, ethylene glycol (EG) and its polymer polyethylene glycol (PEG 400 Da) on the structural and thermal stability of myoglobin (Mb). Our results show that monomer (EG) has no significant effect on the structure of Mb, while the polymer disrupts its structure and decreases its stability. Conversely, the additive effect of crowders showed structural refolding of the protein to some extent. Moreover, the calorimetric binding studies of the protein showed very weak interactions with the mixture of crowders. Usually, we can assume that soft interactions induce structural perturbations while exclusion volume effects stabilize the protein structure; therefore, we hypothesize that under in vivo crowded conditions, both phenomena occur and maintain the stability and function of proteins.https://www.mdpi.com/1420-3049/26/9/2807cellular crowdingsynthetic crowdersmyoglobinprotein stabilityisothermal titration calorimetrymolecular docking |
| spellingShingle | Zahoor Ahmad Parray Faizan Ahmad Md. Imtaiyaz Hassan Anwar Ahmed Fahad N. Almajhdi Ajamaluddin Malik Tajamul Hussain Asimul Islam Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions Molecules cellular crowding synthetic crowders myoglobin protein stability isothermal titration calorimetry molecular docking |
| title | Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions |
| title_full | Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions |
| title_fullStr | Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions |
| title_full_unstemmed | Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions |
| title_short | Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions |
| title_sort | structural refolding and thermal stability of myoglobin in the presence of mixture of crowders importance of various interactions for protein stabilization in crowded conditions |
| topic | cellular crowding synthetic crowders myoglobin protein stability isothermal titration calorimetry molecular docking |
| url | https://www.mdpi.com/1420-3049/26/9/2807 |
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