Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line
Summary: LPR1 (LOW PHOSPHATE ROOT 1), a bacterial-type plant ferroxidase, is crucial for local root phosphate (Pi) sensing. Here, we present a detailed protocol for native (tag-free) protein purification of LPR1 from leaf extracts by differential ammonium sulfate precipitation, size exclusion, and c...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672200613X |
| _version_ | 1797980127252971520 |
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| author | Nancy Tang Christin Naumann |
| author_facet | Nancy Tang Christin Naumann |
| author_sort | Nancy Tang |
| collection | DOAJ |
| description | Summary: LPR1 (LOW PHOSPHATE ROOT 1), a bacterial-type plant ferroxidase, is crucial for local root phosphate (Pi) sensing. Here, we present a detailed protocol for native (tag-free) protein purification of LPR1 from leaf extracts by differential ammonium sulfate precipitation, size exclusion, and cation exchange chromatography of a transgenic Arabidopsis thaliana line overexpressing LPR1. We outline steps for LPR1 purification tracking via immune blot analysis and ferroxidase activity assay. The protocol yields highly pure and active LPR1 protein for biochemical analysis.For complete details on the use and execution of this protocol, please refer to Naumann et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-11T05:50:43Z |
| format | Article |
| id | doaj.art-142f5d383fc8495ea06c37343c9cd24b |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-11T05:50:43Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-142f5d383fc8495ea06c37343c9cd24b2022-12-22T04:42:05ZengElsevierSTAR Protocols2666-16672022-12-0134101733Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana lineNancy Tang0Christin Naumann1Department of Molecular Signal Processing, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120 Halle (Saale), Germany; Corresponding authorDepartment of Molecular Signal Processing, Leibniz Institute of Plant Biochemistry, Weinberg 3, 06120 Halle (Saale), Germany; Corresponding authorSummary: LPR1 (LOW PHOSPHATE ROOT 1), a bacterial-type plant ferroxidase, is crucial for local root phosphate (Pi) sensing. Here, we present a detailed protocol for native (tag-free) protein purification of LPR1 from leaf extracts by differential ammonium sulfate precipitation, size exclusion, and cation exchange chromatography of a transgenic Arabidopsis thaliana line overexpressing LPR1. We outline steps for LPR1 purification tracking via immune blot analysis and ferroxidase activity assay. The protocol yields highly pure and active LPR1 protein for biochemical analysis.For complete details on the use and execution of this protocol, please refer to Naumann 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/S266616672200613XPlant sciencesProtein biochemistryProtein expression and purification |
| spellingShingle | Nancy Tang Christin Naumann Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line STAR Protocols Plant sciences Protein biochemistry Protein expression and purification |
| title | Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line |
| title_full | Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line |
| title_fullStr | Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line |
| title_full_unstemmed | Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line |
| title_short | Native protein purification of ferroxidase LPR1 from leaf extracts of a transgenic Arabidopsis thaliana line |
| title_sort | native protein purification of ferroxidase lpr1 from leaf extracts of a transgenic arabidopsis thaliana line |
| topic | Plant sciences Protein biochemistry Protein expression and purification |
| url | http://www.sciencedirect.com/science/article/pii/S266616672200613X |
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