Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases
Summary: Acyl carrier proteins (ACPs) are key domains in fatty acid and polyketide synthases (PKSs), shuttling intermediates to catalytic partners. Here, we present a protocol for purifying and analyzing ACPs from fatty acid synthases (FASs) and PKSs, using four model ACPs from Mus musculus, Sacchar...
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| Language: | English |
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Elsevier
2025-06-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725001686 |
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| author | Christian Gusenda Svenja Berlage Ines Burkhart Benjamin Chagot Kira J. Weissman Harald Schwalbe Martin Grininger |
| author_facet | Christian Gusenda Svenja Berlage Ines Burkhart Benjamin Chagot Kira J. Weissman Harald Schwalbe Martin Grininger |
| author_sort | Christian Gusenda |
| collection | DOAJ |
| description | Summary: Acyl carrier proteins (ACPs) are key domains in fatty acid and polyketide synthases (PKSs), shuttling intermediates to catalytic partners. Here, we present a protocol for purifying and analyzing ACPs from fatty acid synthases (FASs) and PKSs, using four model ACPs from Mus musculus, Saccharopolyspora erythraea, Streptomyces venezuelae, and Streptomyces hygroscopicus. We describe steps for recombinant ACP production in E. coli; purification via chromatography or precipitation; ACP modification (holo/acyl forms); and analysis using urea PAGE, high-performance liquid chromatography (HPLC), and NMR spectroscopy.For complete details on the use and execution of this protocol, please refer to Gusenda et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-a69b49a58bbb4d67bce878e2fb5636f3 |
| institution | OA Journals |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-a69b49a58bbb4d67bce878e2fb5636f32025-08-20T02:11:50ZengElsevierSTAR Protocols2666-16672025-06-016210376210.1016/j.xpro.2025.103762Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthasesChristian Gusenda0Svenja Berlage1Ines Burkhart2Benjamin Chagot3Kira J. Weissman4Harald Schwalbe5Martin Grininger6Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Buchmann Institute of Molecular Life Sciences (BMLS), Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Corresponding authorInstitute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Buchmann Institute of Molecular Life Sciences (BMLS), Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Université de Lorraine, CNRS, IMoPA, 54000 Nancy, FranceInstitute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Center for Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, 60438 Frankfurt am Main, GermanyUniversité de Lorraine, CNRS, IMoPA, 54000 Nancy, FranceUniversité de Lorraine, CNRS, IMoPA, 54000 Nancy, FranceInstitute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Center for Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, 60438 Frankfurt am Main, GermanyInstitute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Buchmann Institute of Molecular Life Sciences (BMLS), Goethe University Frankfurt, 60438 Frankfurt am Main, Germany; Corresponding authorSummary: Acyl carrier proteins (ACPs) are key domains in fatty acid and polyketide synthases (PKSs), shuttling intermediates to catalytic partners. Here, we present a protocol for purifying and analyzing ACPs from fatty acid synthases (FASs) and PKSs, using four model ACPs from Mus musculus, Saccharopolyspora erythraea, Streptomyces venezuelae, and Streptomyces hygroscopicus. We describe steps for recombinant ACP production in E. coli; purification via chromatography or precipitation; ACP modification (holo/acyl forms); and analysis using urea PAGE, high-performance liquid chromatography (HPLC), and NMR spectroscopy.For complete details on the use and execution of this protocol, please refer to Gusenda et al.1 : 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/S2666166725001686Molecular biologyProtein expression and purificationNMRBiotechnology and bioengineering |
| spellingShingle | Christian Gusenda Svenja Berlage Ines Burkhart Benjamin Chagot Kira J. Weissman Harald Schwalbe Martin Grininger Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases STAR Protocols Molecular biology Protein expression and purification NMR Biotechnology and bioengineering |
| title | Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases |
| title_full | Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases |
| title_fullStr | Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases |
| title_full_unstemmed | Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases |
| title_short | Protocol for the purification, analysis, and handling of acyl carrier proteins from type I fatty acid and polyketide synthases |
| title_sort | protocol for the purification analysis and handling of acyl carrier proteins from type i fatty acid and polyketide synthases |
| topic | Molecular biology Protein expression and purification NMR Biotechnology and bioengineering |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725001686 |
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