Function and maturation of the Fe-S center in dihydroxyacid dehydratase from Arabidopsis
| Autor: | Nicolas Rouhier, Michael K. Johnson, Huanyao Gao, Tamanna Azam, Jérémy Couturier, Sajini Randeniya |
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| Přispěvatelé: | Johnson, Michael K, Dept Chem, University of Adelaide, Ctr Metalloenzyme Studies, Georgia State University, University System of Georgia (USG)-University System of Georgia (USG), Interactions Arbres-Microorganismes (IAM), Université de Lorraine (UL)-Institut National de la Recherche Agronomique (INRA), Novavax, National Institutes of Health R37GM62524, French National Research Agency, 'Investissements d'Avenir' Program ANR-11-LABX-0002-01, Institut National de la Recherche Agronomique (INRA)-Université de Lorraine (UL) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Iron-Sulfur Proteins Circular dichroism [SDV]Life Sciences [q-bio] dihydroxyacid dehydratase Arabidopsis Sequence Homology medicine.disease_cause Spectrum Analysis Raman Biochemistry SCAFFOLD PROTEIN Amino acid synthesis Arabidopsis thaliana NFU protein circular dichroism (CD) electron paramagnetic resonance (EPR) enzyme catalysis iron-sulfur cluster trafficking iron-sulfur protein IRON-SULFUR CLUSTER AZOTOBACTER-VINELANDII (NIF)ISCA MAGNETIC CIRCULAR-DICHROISM SITE-DIRECTED MUTAGENESIS ESCHERICHIA-COLI ACID DEHYDRATASE 2FE-2S CLUSTERS 4FE-4S CLUSTER NITRIC-OXIDE chemistry.chemical_classification biology Circular Dichroism Stereochemistry Iron déshydratase Catalysis Enzyme catalysis 03 medical and health sciences medicine Amino Acid Sequence Molecular Biology Escherichia coli Hydro-Lyases 030102 biochemistry & molecular biology Ligand Arabidopsis Proteins arabidopsis thaliana cluster fe s Cell Biology biology.organism_classification caractérisation in vitro 030104 developmental biology Enzyme chemistry Enzymology Biogenesis Sulfur |
| Zdroj: | Journal of Biological Chemistry 12 (293), 4422-4433. (2018) Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2018, 293 (12), pp.4422-4433. ⟨10.1074/jbc.RA117.001592⟩ |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.RA117.001592⟩ |
| Popis: | Dihydroxyacid dehydratase (DHAD) is the third enzyme required for branched-chain amino acid biosynthesis in bacteria, fungi, and plants. DHAD enzymes contain two distinct types of active-site Fe–S clusters. The best characterized examples are Escherichia coli DHAD, which contains an oxygen-labile [Fe(4)S(4)] cluster, and spinach DHAD, which contains an oxygen-resistant [Fe(2)S(2)] cluster. Although the Fe–S cluster is crucial for DHAD function, little is known about the cluster-coordination environment or the mechanism of catalysis and cluster biogenesis. Here, using the combination of UV-visible absorption and circular dichroism and resonance Raman and electron paramagnetic resonance, we spectroscopically characterized the Fe–S center in DHAD from Arabidopsis thaliana (At). Our results indicated that AtDHAD can accommodate [Fe(2)S(2)] and [Fe(4)S(4)] clusters. However, only the [Fe(2)S(2)] cluster–bound form is catalytically active. We found that the [Fe(2)S(2)] cluster is coordinated by at least one non-cysteinyl ligand, which can be replaced by the thiol group(s) of dithiothreitol. In vitro cluster transfer and reconstitution reactions revealed that [Fe(2)S(2)] cluster–containing NFU2 protein is likely the physiological cluster donor for in vivo maturation of AtDHAD. In summary, AtDHAD binds either one [Fe(4)S(4)] or one [Fe(2)S(2)] cluster, with only the latter being catalytically competent and capable of substrate and product binding, and NFU2 appears to be the physiological [Fe(2)S(2)] cluster donor for DHAD maturation. This work represents the first in vitro characterization of recombinant AtDHAD, providing new insights into the properties, biogenesis, and catalytic role of the active-site Fe–S center in a plant DHAD. |
| Databáze: | OpenAIRE |
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