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Iron Insertion at the Assembly Site of the ISCU Scaffold Protein Is a Conserved Process Initiating Fe-S Cluster Biosynthesis.
Srour, Batoul; Gervason, Sylvain; Hoock, Maren Hellen; Monfort, Beata; Want, Kristian; Larkem, Djabir; Trabelsi, Nadine; Landrot, Gautier; Zitolo, Andrea; Fonda, Emiliano; Etienne, Emilien; Gerbaud, Guillaume; Müller, Christina Sophia; Oltmanns, Jonathan; Gordon, Jesse B; Yadav, Vishal; Kleczewska, Malgorzata; Jelen, Marcin; Toledano, Michel B; Dutkiewicz, Rafal; Goldberg, David P; Schünemann, Volker; Guigliarelli, Bruno; Burlat, Bénédicte; Sizun, Christina; D'Autréaux, Benoit.
Afiliación
  • Srour B; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Gervason S; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Hoock MH; Fachbereich Physik, Technische Universität Kaiserslautern, Erwin-Schrödinger-Str. 56, 67663 Kaiserslautern, Germany.
  • Monfort B; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Want K; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Larkem D; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Trabelsi N; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Landrot G; Synchrotron SOLEIL, L'Orme des Merisiers, BP48 Saint Aubin 91192 Gif-Sur-Yvette, France.
  • Zitolo A; Synchrotron SOLEIL, L'Orme des Merisiers, BP48 Saint Aubin 91192 Gif-Sur-Yvette, France.
  • Fonda E; Synchrotron SOLEIL, L'Orme des Merisiers, BP48 Saint Aubin 91192 Gif-Sur-Yvette, France.
  • Etienne E; Aix Marseille Univ, CNRS, Laboratoire de Bioénergétique et Ingénierie des Protéines (BIP), 31 Chemin Joseph Aiguier, 13402 Marseille, France.
  • Gerbaud G; Aix Marseille Univ, CNRS, Laboratoire de Bioénergétique et Ingénierie des Protéines (BIP), 31 Chemin Joseph Aiguier, 13402 Marseille, France.
  • Müller CS; Fachbereich Physik, Technische Universität Kaiserslautern, Erwin-Schrödinger-Str. 56, 67663 Kaiserslautern, Germany.
  • Oltmanns J; Fachbereich Physik, Technische Universität Kaiserslautern, Erwin-Schrödinger-Str. 56, 67663 Kaiserslautern, Germany.
  • Gordon JB; Department of Chemistry, The Johns Hopkins University, 3400 N. Charles Street, Baltimore, Maryland 21218, United States.
  • Yadav V; Department of Chemistry, The Johns Hopkins University, 3400 N. Charles Street, Baltimore, Maryland 21218, United States.
  • Kleczewska M; Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Abrahama 58, 80-307 Gdansk, Poland.
  • Jelen M; Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Abrahama 58, 80-307 Gdansk, Poland.
  • Toledano MB; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
  • Dutkiewicz R; Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Abrahama 58, 80-307 Gdansk, Poland.
  • Goldberg DP; Department of Chemistry, The Johns Hopkins University, 3400 N. Charles Street, Baltimore, Maryland 21218, United States.
  • Schünemann V; Fachbereich Physik, Technische Universität Kaiserslautern, Erwin-Schrödinger-Str. 56, 67663 Kaiserslautern, Germany.
  • Guigliarelli B; Aix Marseille Univ, CNRS, Laboratoire de Bioénergétique et Ingénierie des Protéines (BIP), 31 Chemin Joseph Aiguier, 13402 Marseille, France.
  • Burlat B; Aix Marseille Univ, CNRS, Laboratoire de Bioénergétique et Ingénierie des Protéines (BIP), 31 Chemin Joseph Aiguier, 13402 Marseille, France.
  • Sizun C; Institut de Chimie des Substances Naturelles, CNRS, Université Paris Saclay, Avenue de La Terrasse, 91190 Gif-sur-Yvette, France.
  • D'Autréaux B; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198 Gif-sur-Yvette, France.
J Am Chem Soc ; 144(38): 17496-17515, 2022 09 28.
Article en En | MEDLINE | ID: mdl-36121382
Iron-sulfur (Fe-S) clusters are prosthetic groups of proteins biosynthesized on scaffold proteins by highly conserved multi-protein machineries. Biosynthesis of Fe-S clusters into the ISCU scaffold protein is initiated by ferrous iron insertion, followed by sulfur acquisition, via a still elusive mechanism. Notably, whether iron initially binds to the ISCU cysteine-rich assembly site or to a cysteine-less auxiliary site via N/O ligands remains unclear. We show here by SEC, circular dichroism (CD), and Mössbauer spectroscopies that iron binds to the assembly site of the monomeric form of prokaryotic and eukaryotic ISCU proteins via either one or two cysteines, referred to the 1-Cys and 2-Cys forms, respectively. The latter predominated at pH 8.0 and correlated with the Fe-S cluster assembly activity, whereas the former increased at a more acidic pH, together with free iron, suggesting that it constitutes an intermediate of the iron insertion process. Iron not binding to the assembly site was non-specifically bound to the aggregated ISCU, ruling out the existence of a structurally defined auxiliary site in ISCU. Characterization of the 2-Cys form by site-directed mutagenesis, CD, NMR, X-ray absorption, Mössbauer, and electron paramagnetic resonance spectroscopies showed that the iron center is coordinated by four strictly conserved amino acids of the assembly site, Cys35, Asp37, Cys61, and His103, in a tetrahedral geometry. The sulfur receptor Cys104 was at a very close distance and apparently bound to the iron center when His103 was missing, which may enable iron-dependent sulfur acquisition. Altogether, these data provide the structural basis to elucidate the Fe-S cluster assembly process and establish that the initiation of Fe-S cluster biosynthesis by insertion of a ferrous iron in the assembly site of ISCU is a conserved mechanism.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Escherichia coli / Proteínas Hierro-Azufre Idioma: En Revista: J Am Chem Soc Año: 2022 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Estados Unidos
Texto completo
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Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Escherichia coli / Proteínas Hierro-Azufre Idioma: En Revista: J Am Chem Soc Año: 2022 Tipo del documento: Article País de afiliación: Francia Pais de publicación: Estados Unidos