Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis
Summary: We present an in vitro and in-cell activity-based protein profiling (ABPP) protocol for endogenous nonribosomal peptide synthetases (NRPSs). This protocol enables the fluorescence labeling and imaging of an endogenous SrfAB-NRPS with high selectivity and sensitivity in the surfactin produce...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-09-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722003422 |
| _version_ | 1811243365386158080 |
|---|---|
| author | Fumihiro Ishikawa Rina Ohnishi Chiharu Uchida Genzoh Tanabe |
| author_facet | Fumihiro Ishikawa Rina Ohnishi Chiharu Uchida Genzoh Tanabe |
| author_sort | Fumihiro Ishikawa |
| collection | DOAJ |
| description | Summary: We present an in vitro and in-cell activity-based protein profiling (ABPP) protocol for endogenous nonribosomal peptide synthetases (NRPSs). This protocol enables the fluorescence labeling and imaging of an endogenous SrfAB-NRPS with high selectivity and sensitivity in the surfactin producer Bacillus subtilis. While we optimized this protocol for use with B. subtilis, the protocol can be applied to Aneurinibacillus migulanus and Escherichia coli.For complete details on the use and execution of this protocol, please refer to Ishikawa et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-12T14:06:10Z |
| format | Article |
| id | doaj.art-2b58ac9f0ef6490385c0e9d297ca1eb7 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-12T14:06:10Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-2b58ac9f0ef6490385c0e9d297ca1eb72022-12-22T03:30:03ZengElsevierSTAR Protocols2666-16672022-09-0133101462Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilisFumihiro Ishikawa0Rina Ohnishi1Chiharu Uchida2Genzoh Tanabe3Faculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan; Corresponding authorFaculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, JapanFaculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, JapanFaculty of Pharmacy, Kindai University, 3-4-1 Kowakae, Higashi-Osaka, Osaka 577-8502, Japan; Corresponding authorSummary: We present an in vitro and in-cell activity-based protein profiling (ABPP) protocol for endogenous nonribosomal peptide synthetases (NRPSs). This protocol enables the fluorescence labeling and imaging of an endogenous SrfAB-NRPS with high selectivity and sensitivity in the surfactin producer Bacillus subtilis. While we optimized this protocol for use with B. subtilis, the protocol can be applied to Aneurinibacillus migulanus and Escherichia coli.For complete details on the use and execution of this protocol, please refer to Ishikawa et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722003422MicrobiologyMolecular/Chemical ProbesProtein BiochemistryChemistry |
| spellingShingle | Fumihiro Ishikawa Rina Ohnishi Chiharu Uchida Genzoh Tanabe Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis STAR Protocols Microbiology Molecular/Chemical Probes Protein Biochemistry Chemistry |
| title | Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis |
| title_full | Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis |
| title_fullStr | Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis |
| title_full_unstemmed | Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis |
| title_short | Activity-based protein profiling of a surfactin-producing nonribosomal peptide synthetase in Bacillus subtilis |
| title_sort | activity based protein profiling of a surfactin producing nonribosomal peptide synthetase in bacillus subtilis |
| topic | Microbiology Molecular/Chemical Probes Protein Biochemistry Chemistry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722003422 |
| work_keys_str_mv | AT fumihiroishikawa activitybasedproteinprofilingofasurfactinproducingnonribosomalpeptidesynthetaseinbacillussubtilis AT rinaohnishi activitybasedproteinprofilingofasurfactinproducingnonribosomalpeptidesynthetaseinbacillussubtilis AT chiharuuchida activitybasedproteinprofilingofasurfactinproducingnonribosomalpeptidesynthetaseinbacillussubtilis AT genzohtanabe activitybasedproteinprofilingofasurfactinproducingnonribosomalpeptidesynthetaseinbacillussubtilis |