دورية أكاديمية
The A-type domain in Escherichia coli NfuA is required for regenerating the auxiliary [4Fe-4S] cluster in Escherichia coli lipoyl synthase.
| العنوان: | The A-type domain in Escherichia coli NfuA is required for regenerating the auxiliary [4Fe-4S] cluster in Escherichia coli lipoyl synthase. |
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| المؤلفون: | McCarthy EL; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802., Rankin AN; Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802., Dill ZR; Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802., Booker SJ; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, Pennsylvania 16802; Department of Chemistry, The Pennsylvania State University, University Park, Pennsylvania 16802; Howard Hughes Medical Institute, The Pennsylvania State University, University Park, Pennsylvania 16802. Electronic address: squire@psu.edu. |
| المصدر: | The Journal of biological chemistry [J Biol Chem] 2019 Feb 01; Vol. 294 (5), pp. 1609-1617. Date of Electronic Publication: 2018 Dec 11. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology |
| مواضيع طبية MeSH: | Bacterial Proteins/*metabolism , Escherichia coli/*enzymology , Escherichia coli Proteins/*metabolism , Iron/*metabolism , Iron-Sulfur Proteins/*metabolism , Sulfur/*metabolism, Amino Acid Sequence ; Bacterial Proteins/chemistry ; Catalysis ; Escherichia coli Proteins/chemistry ; Iron/chemistry ; Iron-Sulfur Proteins/chemistry ; Protein Domains ; Sulfur/chemistry |
| مستخلص: | The lipoyl cofactor plays an integral role in several essential biological processes. The last step in its de novo biosynthetic pathway, the attachment of two sulfur atoms at C6 and C8 of an n -octanoyllysyl chain, is catalyzed by lipoyl synthase (LipA), a member of the radical SAM superfamily. In addition to the [4Fe-4S] cluster common to all radical SAM enzymes, LipA contains a second [4Fe-4S] auxiliary cluster, which is sacrificed during catalysis to supply the requisite sulfur atoms, rendering the protein inactive for further turnovers. Recently, it was shown that the Fe-S cluster carrier protein NfuA from Escherichia coli can regenerate the auxiliary cluster of E. coli LipA after each turnover, but the molecular mechanism is incompletely understood. Herein, using protein-protein interaction and kinetic assays as well as site-directed mutagenesis, we provide further insight into the mechanism of NfuA-mediated cluster regeneration. In particular, we show that the N-terminal A-type domain of E. coli NfuA is essential for its tight interaction with LipA. Further, we demonstrate that NfuA from Mycobacterium tuberculosis can also regenerate the auxiliary cluster of E. coli LipA. However, an Nfu protein from Staphylococcus aureus , which lacks the A-type domain, was severely diminished in facilitating cluster regeneration. Of note, addition of the N-terminal domain of E. coli NfuA to S. aureus Nfu, fully restored cluster-regenerating activity. These results expand our understanding of the newly discovered mechanism by which the auxiliary cluster of LipA is restored after each turnover. (© 2019 McCarthy et al.) |
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| معلومات مُعتمدة: | R35 GM122595 United States GM NIGMS NIH HHS; United States HHMI Howard Hughes Medical Institute |
| فهرسة مساهمة: | Keywords: Fe-S cluster carrier; NfuA; S-adenosylmethionine (SAM); enzyme mechanism; iron-sulfur protein; lipoic acid; metalloenzyme; radical; radical SAM enzyme; sulfur |
| المشرفين على المادة: | 0 (Bacterial Proteins) 0 (Escherichia coli Proteins) 0 (Iron-Sulfur Proteins) 0 (LipA protein, Bacteria) 0 (NfuA protein, E coli) 70FD1KFU70 (Sulfur) E1UOL152H7 (Iron) |
| تواريخ الأحداث: | Date Created: 20181213 Date Completed: 20190520 Latest Revision: 20210314 |
| رمز التحديث: | 20240628 |
| مُعرف محوري في PubMed: | PMC6364782 |
| DOI: | 10.1074/jbc.RA118.006171 |
| PMID: | 30538130 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1083-351X |
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| DOI: | 10.1074/jbc.RA118.006171 |