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Peroxynitrite and nitric oxide differently target the iron-sulfur cluster and amino acid residues of human iron regulatory protein 1.
Biochemistry. 2003 Jul 01; 42(25):7648-54.B

Abstract

Iron regulatory protein 1 (IRP1) is a redox-sensitive protein which exists in two active forms in the cytosol of eukaryotic cells. Holo-IRP1 containing a [4Fe-4S] cluster exhibits aconitase activity which catalyzes the isomerization of citrate and isocitrate. The cluster-free protein (apo-IRP1) is a transregulator binding to specific mRNA, and thus post-transcriptionally modulating the expression of genes involved in iron metabolism. The resonance Raman (RR) spectra of human recombinant holo-IRP1 (rhIRP1) excited at 457.9 nm show that the 395 cm(-1) band, attributed to a terminal Fe-S stretching mode of the cluster, is replaced by a 405 cm(-1) band, consistent with the conversion of the [4Fe-4S](2+) center to a [3Fe-4S](+) center, upon exposure to peroxynitrite. This conclusion was confirmed by electron paramagnetic resonance (EPR) data and correlated with the loss of aconitase activity. In another series of experiments, the RR spectra also revealed the presence of additional bands at 818 and 399 cm(-1) when rhIRP1 was treated with a peroxynitrite synthesized by a different procedure. These bands correspond to those of 3-nitrotyrosine, and they indicate nitration of at least one tyrosine residue in rhIRP1. This was further confirmed by Western blot analysis with an anti-nitrotyrosine antibody. In contrast, the reaction of rhIRP1 with NO in the absence of oxygen revealed full mRNA binding activity of the protein, without nitration of tyrosines. These results strongly suggest that NO mainly acts as a regulator of IRP1 whereas peroxynitrite is likely to disrupt the IRP1/IRE regulatory pathway.

Authors+Show Affiliations

Institut de Chimie des Substances Naturelles, CNRS, 91190 Gif-sur-Yvette, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12820873

Citation

Soum, Emmanuelle, et al. "Peroxynitrite and Nitric Oxide Differently Target the Iron-sulfur Cluster and Amino Acid Residues of Human Iron Regulatory Protein 1." Biochemistry, vol. 42, no. 25, 2003, pp. 7648-54.
Soum E, Brazzolotto X, Goussias C, et al. Peroxynitrite and nitric oxide differently target the iron-sulfur cluster and amino acid residues of human iron regulatory protein 1. Biochemistry. 2003;42(25):7648-54.
Soum, E., Brazzolotto, X., Goussias, C., Bouton, C., Moulis, J. M., Mattioli, T. A., & Drapier, J. C. (2003). Peroxynitrite and nitric oxide differently target the iron-sulfur cluster and amino acid residues of human iron regulatory protein 1. Biochemistry, 42(25), 7648-54.
Soum E, et al. Peroxynitrite and Nitric Oxide Differently Target the Iron-sulfur Cluster and Amino Acid Residues of Human Iron Regulatory Protein 1. Biochemistry. 2003 Jul 1;42(25):7648-54. PubMed PMID: 12820873.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Peroxynitrite and nitric oxide differently target the iron-sulfur cluster and amino acid residues of human iron regulatory protein 1. AU - Soum,Emmanuelle, AU - Brazzolotto,Xavier, AU - Goussias,Charilaos, AU - Bouton,Cécile, AU - Moulis,Jean-Marc, AU - Mattioli,Tony A, AU - Drapier,Jean-Claude, PY - 2003/6/25/pubmed PY - 2003/8/27/medline PY - 2003/6/25/entrez SP - 7648 EP - 54 JF - Biochemistry JO - Biochemistry VL - 42 IS - 25 N2 - Iron regulatory protein 1 (IRP1) is a redox-sensitive protein which exists in two active forms in the cytosol of eukaryotic cells. Holo-IRP1 containing a [4Fe-4S] cluster exhibits aconitase activity which catalyzes the isomerization of citrate and isocitrate. The cluster-free protein (apo-IRP1) is a transregulator binding to specific mRNA, and thus post-transcriptionally modulating the expression of genes involved in iron metabolism. The resonance Raman (RR) spectra of human recombinant holo-IRP1 (rhIRP1) excited at 457.9 nm show that the 395 cm(-1) band, attributed to a terminal Fe-S stretching mode of the cluster, is replaced by a 405 cm(-1) band, consistent with the conversion of the [4Fe-4S](2+) center to a [3Fe-4S](+) center, upon exposure to peroxynitrite. This conclusion was confirmed by electron paramagnetic resonance (EPR) data and correlated with the loss of aconitase activity. In another series of experiments, the RR spectra also revealed the presence of additional bands at 818 and 399 cm(-1) when rhIRP1 was treated with a peroxynitrite synthesized by a different procedure. These bands correspond to those of 3-nitrotyrosine, and they indicate nitration of at least one tyrosine residue in rhIRP1. This was further confirmed by Western blot analysis with an anti-nitrotyrosine antibody. In contrast, the reaction of rhIRP1 with NO in the absence of oxygen revealed full mRNA binding activity of the protein, without nitration of tyrosines. These results strongly suggest that NO mainly acts as a regulator of IRP1 whereas peroxynitrite is likely to disrupt the IRP1/IRE regulatory pathway. SN - 0006-2960 UR - https://www.unboundmedicine.com/medline/citation/12820873/Peroxynitrite_and_nitric_oxide_differently_target_the_iron_sulfur_cluster_and_amino_acid_residues_of_human_iron_regulatory_protein_1_ L2 - https://doi.org/10.1021/bi030041i DB - PRIME DP - Unbound Medicine ER -