دورية أكاديمية
Cytochrome c nitrite reductase from the bacterium Geobacter lovleyi represents a new NrfA subclass.
| العنوان: | Cytochrome c nitrite reductase from the bacterium Geobacter lovleyi represents a new NrfA subclass. |
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| المؤلفون: | Campeciño J; Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, USA., Lagishetty S; Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, USA., Wawrzak Z; Synchrotron Research Center, Life Science Collaborative Access Team, Northwestern University, Argonne, Illinois, USA., Sosa Alfaro V; Department of Chemistry and Biophysics, The University of Michigan, Ann Arbor, Michigan, USA., Lehnert N; Department of Chemistry and Biophysics, The University of Michigan, Ann Arbor, Michigan, USA., Reguera G; Department of Microbiology & Molecular Genetics, Michigan State University, East Lansing, Michigan, USA., Hu J; Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, USA.; Department of Chemistry, Michigan State University, East Lansing, Michigan, USA., Hegg EL; Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, USA EricHegg@msu.edu. |
| المصدر: | The Journal of biological chemistry [J Biol Chem] 2020 Aug 14; Vol. 295 (33), pp. 11455-11465. Date of Electronic Publication: 2020 Jun 09. |
| نوع المنشور: | Journal Article; 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 , Cytochromes a1/*metabolism , Cytochromes c1/*metabolism , Geobacter/*metabolism , Nitrate Reductases/*metabolism, Ammonium Compounds/metabolism ; Bacterial Proteins/chemistry ; Bacterial Proteins/genetics ; Crystallography, X-Ray ; Cytochromes a1/chemistry ; Cytochromes a1/genetics ; Cytochromes c1/chemistry ; Cytochromes c1/genetics ; Geobacter/chemistry ; Geobacter/genetics ; Kinetics ; Models, Molecular ; Nitrate Reductases/chemistry ; Nitrate Reductases/genetics ; Nitrates/metabolism ; Phylogeny ; Protein Conformation |
| مستخلص: | Cytochrome c nitrite reductase (NrfA) catalyzes the reduction of nitrite to ammonium in the dissimilatory nitrate reduction to ammonium (DNRA) pathway, a process that competes with denitrification, conserves nitrogen, and minimizes nutrient loss in soils. The environmental bacterium Geobacter lovleyi has recently been recognized as a key driver of DNRA in nature, but its enzymatic pathway is still uncharacterized. To address this limitation, here we overexpressed, purified, and characterized G. lovleyi NrfA. We observed that the enzyme crystallizes as a dimer but remains monomeric in solution. Importantly, its crystal structure at 2.55-Å resolution revealed the presence of an arginine residue in the region otherwise occupied by calcium in canonical NrfA enzymes. The presence of EDTA did not affect the activity of G. lovleyi NrfA, and site-directed mutagenesis of this arginine reduced enzymatic activity to <3% of the WT levels. Phylogenetic analysis revealed four separate emergences of Arg-containing NrfA enzymes. Thus, the Ca 2+ -independent, Arg-containing NrfA from G. lovleyi represents a new subclass of cytochrome c nitrite reductase. Most genera from the exclusive clades of Arg-containing NrfA proteins are also represented in clades containing Ca 2+ -dependent enzymes, suggesting convergent evolution. Competing Interests: Conflict of interest—The authors declare no conflicts of interest in regards to this work. (© 2020 Campeciño et al.) |
| التعليقات: | Erratum in: J Biol Chem. 2022 Dec;298(12):102743. (PMID: 36473407) |
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| فهرسة مساهمة: | Keywords: Geobacter lovleyi; NrfA; calcium; crystal structure; cytochrome c nitrite reductase; dissimilatory nitrate reduction to ammonium (DNRA); enzyme catalysis; metalloenzyme; nitrogen cycle; phylogenetics |
| سلسلة جزيئية: | PDB 1QDB |
| المشرفين على المادة: | 0 (Ammonium Compounds) 0 (Bacterial Proteins) 0 (Nitrates) 9035-35-2 (Cytochromes a1) 9035-42-1 (Cytochromes c1) EC 1.7.- (Nitrate Reductases) EC 1.9.6.1 (nitrate reductase (cytochrome)) |
| SCR Organism: | Geobacter lovleyi |
| تواريخ الأحداث: | Date Created: 20200611 Date Completed: 20210119 Latest Revision: 20230106 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC7450111 |
| DOI: | 10.1074/jbc.RA120.013981 |
| PMID: | 32518164 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1083-351X |
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| DOI: | 10.1074/jbc.RA120.013981 |