Structural investigation of APRs to improve the solubility of outer membrane protease (PgtE) from Salmonella enterica serotype typhi- A multi-constraint approach
| Autor: | Sundarabaalaji Narayanan, Christian Bharathi Antonyraj, Gopinath Samykannu, Princy Vijayababu |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Salmonella enteric serotype typhi medicine.medical_treatment Omptin PgtE Biophysics Biochemistry lcsh:Biochemistry 03 medical and health sciences 0302 clinical medicine Protein structure Aspartic acid Solubilization medicine CABS flex lcsh:QD415-436 Denaturation (biochemistry) lcsh:QH301-705.5 Protease biology Chemistry APRs biology.organism_classification Chaotropic agent 030104 developmental biology lcsh:Biology (General) Salmonella enterica 030220 oncology & carcinogenesis Bacterial outer membrane Structural gatekeepers Research Article |
| Zdroj: | Biochemistry and Biophysics Reports Biochemistry and Biophysics Reports, Vol 21, Iss, Pp-(2020) |
| ISSN: | 2405-5808 |
| DOI: | 10.1016/j.bbrep.2019.100693 |
| Popis: | Outer membrane proteins were playing a crucial role on the several functions controlled by cell membranes even though they are not naturally expressed at higher levels. In order to obtain biologically active protein, the denaturation of these inclusion bodies must be optimized using chaotropic agents. Hence, this study focuses on improving the yield of Outer Membrane Protease (PgtE) from Salmonella enterica serotype Typhi (S. Typhi) using chaotropes and additives. Denaturation methods were tried with various pH, detergents, and reducing agents were used to optimize the solubility of PgtE with biologically active form. Due to the aggregation, we failed to achieve the maximum yield of PgtE. Consequently, we predicted 9 Aggregation Prone Regions (APRs) in PgtE, which are mutated by known structural Gatekeepers. We calculated the Aggregation Index (AI) of PgtE with 10 mM of aspartic acid as an additive in optimized buffer. In addition, the mutations at specific positions within the protein structure can act as APRs suppressors without affecting protein stability with CABS flex dynamics. The multiple sequence analysis demonstrate that aspartic acid is appropriate denaturing additive for other Gram-negative pathogens of Omptin family. Graphical abstract Image 1 |
| Databáze: | OpenAIRE |
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