Complementary protocols to evaluate inhibitors against the DnaK chaperone network
Summary: Bacterial DnaK belongs to the Hsp70 chaperone family, which plays a critical role in maintaining proteostasis by catalyzing protein folding, and is a proposed antibacterial target in the pathogen Mycobacterium tuberculosis. Here, we describe an experimental toolbox for evaluating inhibitors...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722002611 |
| _version_ | 1818005986930065408 |
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| author | Aweon Richards Gideon K. Yawson Brock Nelson Tania J. Lupoli |
| author_facet | Aweon Richards Gideon K. Yawson Brock Nelson Tania J. Lupoli |
| author_sort | Aweon Richards |
| collection | DOAJ |
| description | Summary: Bacterial DnaK belongs to the Hsp70 chaperone family, which plays a critical role in maintaining proteostasis by catalyzing protein folding, and is a proposed antibacterial target in the pathogen Mycobacterium tuberculosis. Here, we describe an experimental toolbox for evaluating inhibitors against the mycobacterial DnaK chaperone network: a coupled-enzymatic assay to monitor ATPase activity, a proteolytic cleavage assay to study DnaK conformational changes upon ligand addition, as well as a protein renaturation assay to assess chaperone function.For complete details on the use and execution of this protocol, please refer to Hosfelt et al. (2021). |
| first_indexed | 2024-04-14T04:54:18Z |
| format | Article |
| id | doaj.art-d32f51e1d03f4b659ca5fb3ec331492e |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-14T04:54:18Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-d32f51e1d03f4b659ca5fb3ec331492e2022-12-22T02:11:12ZengElsevierSTAR Protocols2666-16672022-06-0132101381Complementary protocols to evaluate inhibitors against the DnaK chaperone networkAweon Richards0Gideon K. Yawson1Brock Nelson2Tania J. Lupoli3Department of Chemistry, New York University, New York, NY 10003, USADepartment of Chemistry, New York University, New York, NY 10003, USADepartment of Chemistry, New York University, New York, NY 10003, USADepartment of Chemistry, New York University, New York, NY 10003, USA; Corresponding authorSummary: Bacterial DnaK belongs to the Hsp70 chaperone family, which plays a critical role in maintaining proteostasis by catalyzing protein folding, and is a proposed antibacterial target in the pathogen Mycobacterium tuberculosis. Here, we describe an experimental toolbox for evaluating inhibitors against the mycobacterial DnaK chaperone network: a coupled-enzymatic assay to monitor ATPase activity, a proteolytic cleavage assay to study DnaK conformational changes upon ligand addition, as well as a protein renaturation assay to assess chaperone function.For complete details on the use and execution of this protocol, please refer to Hosfelt et al. (2021).http://www.sciencedirect.com/science/article/pii/S2666166722002611Molecular/Chemical ProbesProtein Biochemistry |
| spellingShingle | Aweon Richards Gideon K. Yawson Brock Nelson Tania J. Lupoli Complementary protocols to evaluate inhibitors against the DnaK chaperone network STAR Protocols Molecular/Chemical Probes Protein Biochemistry |
| title | Complementary protocols to evaluate inhibitors against the DnaK chaperone network |
| title_full | Complementary protocols to evaluate inhibitors against the DnaK chaperone network |
| title_fullStr | Complementary protocols to evaluate inhibitors against the DnaK chaperone network |
| title_full_unstemmed | Complementary protocols to evaluate inhibitors against the DnaK chaperone network |
| title_short | Complementary protocols to evaluate inhibitors against the DnaK chaperone network |
| title_sort | complementary protocols to evaluate inhibitors against the dnak chaperone network |
| topic | Molecular/Chemical Probes Protein Biochemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722002611 |
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