دورية أكاديمية
The unique Phe-His dyad of 2-ketopropyl coenzyme M oxidoreductase/carboxylase selectively promotes carboxylation and S-C bond cleavage.
| العنوان: | The unique Phe-His dyad of 2-ketopropyl coenzyme M oxidoreductase/carboxylase selectively promotes carboxylation and S-C bond cleavage. |
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| المؤلفون: | Prussia GA; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA., Shisler KA; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA., Zadvornyy OA; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA., Streit BR; Department of Chemistry and Biochemistry, Montana State University, Bozeman, Montana, USA., DuBois JL; Department of Chemistry and Biochemistry, Montana State University, Bozeman, Montana, USA., Peters JW; Institute of Biological Chemistry, Washington State University, Pullman, Washington, USA. Electronic address: jw.peters@wsu.edu. |
| المصدر: | The Journal of biological chemistry [J Biol Chem] 2021 Aug; Vol. 297 (2), pp. 100961. Date of Electronic Publication: 2021 Jul 12. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; 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 |
| 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: | Histidine*/chemistry , Histidine*/metabolism, Phenylalanine/chemistry ; Phenylalanine/metabolism ; Crystallography, X-Ray ; Acetoacetates/chemistry ; Acetoacetates/metabolism ; Amino Acid Substitution ; Ketone Oxidoreductases/metabolism ; Ketone Oxidoreductases/chemistry |
| مستخلص: | The 2-ketopropyl-coenzyme M oxidoreductase/carboxylase (2-KPCC) enzyme is the only member of the disulfide oxidoreductase (DSOR) family of enzymes, which are important for reductively cleaving S-S bonds, to have carboxylation activity. 2-KPCC catalyzes the conversion of 2-ketopropyl-coenzyme M to acetoacetate, which is used as a carbon source, in a controlled reaction to exclude protons. A conserved His-Glu motif present in DSORs is key in the protonation step; however, in 2-KPCC, the dyad is substituted by Phe-His. Here, we propose that this difference is important for coupling carboxylation with C-S bond cleavage. We substituted the Phe-His dyad in 2-KPCC to be more DSOR like, replacing the phenylalanine with histidine (F501H) and the histidine with glutamate (H506E), and solved crystal structures of F501H and the double variant F501H_H506E. We found that F501 protects the enolacetone intermediate from protons and that the F501H variant strongly promotes protonation. We also provided evidence for the involvement of the H506 residue in stabilizing the developing charge during the formation of acetoacetate, which acts as a product inhibitor in the WT but not the H506E variant enzymes. Finally, we determined that the F501H substitution promotes a DSOR-like charge transfer interaction with flavin adenine dinucleotide, eliminating the need for cysteine as an internal base. Taken together, these results indicate that the 2-KPCC dyad is responsible for selectively promoting carboxylation and inhibiting protonation in the formation of acetoacetate. Competing Interests: Conflict of interest The authors declare that they have no conflict of interest with the contents of this article. (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.) |
| معلومات مُعتمدة: | P41 GM103393 United States GM NIGMS NIH HHS |
| فهرسة مساهمة: | Keywords: anion-binding; carbon dioxide fixation; carbon–sulfur bond; carboxylation; kinetics; oxidation–reduction |
| المشرفين على المادة: | 4QD397987E (Histidine) 47E5O17Y3R (Phenylalanine) 0 (Acetoacetates) 4ZI204Y1MC (acetoacetic acid) EC 1.2.- (Ketone Oxidoreductases) |
| تواريخ الأحداث: | Date Created: 20210715 Date Completed: 20240725 Latest Revision: 20240729 |
| رمز التحديث: | 20240729 |
| مُعرف محوري في PubMed: | PMC8358701 |
| DOI: | 10.1016/j.jbc.2021.100961 |
| PMID: | 34265301 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1083-351X |
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| DOI: | 10.1016/j.jbc.2021.100961 |