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New Insights into Hemopexin-Binding to Hemin and Hemoglobin

Hemopexin (Hx) is a plasma glycoprotein that scavenges heme (Fe(III) protoporphyrin IX). Hx has important implications in hemolytic disorders and hemorrhagic conditions because releasing hemoglobin increases the labile heme, which is potentially toxic, thus producing oxidative stress. Therefore, Hx...

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Autores principales: Lechuga, Guilherme C., Napoleão-Pêgo, Paloma, Morel, Carlos M., Provance, David W., De-Simone, Salvatore G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8998376/
https://www.ncbi.nlm.nih.gov/pubmed/35409149
http://dx.doi.org/10.3390/ijms23073789
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author Lechuga, Guilherme C.
Napoleão-Pêgo, Paloma
Morel, Carlos M.
Provance, David W.
De-Simone, Salvatore G.
author_facet Lechuga, Guilherme C.
Napoleão-Pêgo, Paloma
Morel, Carlos M.
Provance, David W.
De-Simone, Salvatore G.
author_sort Lechuga, Guilherme C.
collection PubMed
description Hemopexin (Hx) is a plasma glycoprotein that scavenges heme (Fe(III) protoporphyrin IX). Hx has important implications in hemolytic disorders and hemorrhagic conditions because releasing hemoglobin increases the labile heme, which is potentially toxic, thus producing oxidative stress. Therefore, Hx has been considered for therapeutic use and diagnostics. In this work, we analyzed and mapped the interaction sequences of Hx with hemin and hemoglobin. The spot-synthesis technique was used to map human hemopexin (P02790) binding to hemin and human hemoglobin. A library of 15 amino acid peptides with a 10-amino acid overlap was designed to represent the entire coding region (aa 1-462) of hemopexin and synthesized onto cellulose membranes. An in silico approach was taken to analyze the amino acid frequency in the identified interaction regions, and molecular docking was applied to assess the protein-protein interaction. Seven linear peptide sequences in Hx were identified to bind hemin (H1–H7), and five were described for Hb (Hb1–Hb5) interaction, with just two sequences shared between hemin and Hb. The amino acid composition of the identified sequences demonstrated that histidine residues are relevant for heme binding. H105, H293, H373, H400, H429, and H462 were distributed in the H1–H7 peptide sequences, but other residues may also play an important role. Molecular docking analysis demonstrated Hx’s association with the β-chain of Hb, with several hotspot amino acids that coordinated the interaction. This study provides new insights into Hx-hemin binding motifs and protein-protein interactions with Hb. The identified binding sequences and specific peptides can be used for therapeutic purposes and diagnostics as hemopexin is under investigation to treat different diseases and there is an urgent need for diagnostics using labile heme when monitoring hemolysis.
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spelling pubmed-89983762022-04-12 New Insights into Hemopexin-Binding to Hemin and Hemoglobin Lechuga, Guilherme C. Napoleão-Pêgo, Paloma Morel, Carlos M. Provance, David W. De-Simone, Salvatore G. Int J Mol Sci Article Hemopexin (Hx) is a plasma glycoprotein that scavenges heme (Fe(III) protoporphyrin IX). Hx has important implications in hemolytic disorders and hemorrhagic conditions because releasing hemoglobin increases the labile heme, which is potentially toxic, thus producing oxidative stress. Therefore, Hx has been considered for therapeutic use and diagnostics. In this work, we analyzed and mapped the interaction sequences of Hx with hemin and hemoglobin. The spot-synthesis technique was used to map human hemopexin (P02790) binding to hemin and human hemoglobin. A library of 15 amino acid peptides with a 10-amino acid overlap was designed to represent the entire coding region (aa 1-462) of hemopexin and synthesized onto cellulose membranes. An in silico approach was taken to analyze the amino acid frequency in the identified interaction regions, and molecular docking was applied to assess the protein-protein interaction. Seven linear peptide sequences in Hx were identified to bind hemin (H1–H7), and five were described for Hb (Hb1–Hb5) interaction, with just two sequences shared between hemin and Hb. The amino acid composition of the identified sequences demonstrated that histidine residues are relevant for heme binding. H105, H293, H373, H400, H429, and H462 were distributed in the H1–H7 peptide sequences, but other residues may also play an important role. Molecular docking analysis demonstrated Hx’s association with the β-chain of Hb, with several hotspot amino acids that coordinated the interaction. This study provides new insights into Hx-hemin binding motifs and protein-protein interactions with Hb. The identified binding sequences and specific peptides can be used for therapeutic purposes and diagnostics as hemopexin is under investigation to treat different diseases and there is an urgent need for diagnostics using labile heme when monitoring hemolysis. MDPI 2022-03-30 /pmc/articles/PMC8998376/ /pubmed/35409149 http://dx.doi.org/10.3390/ijms23073789 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lechuga, Guilherme C.
Napoleão-Pêgo, Paloma
Morel, Carlos M.
Provance, David W.
De-Simone, Salvatore G.
New Insights into Hemopexin-Binding to Hemin and Hemoglobin
title New Insights into Hemopexin-Binding to Hemin and Hemoglobin
title_full New Insights into Hemopexin-Binding to Hemin and Hemoglobin
title_fullStr New Insights into Hemopexin-Binding to Hemin and Hemoglobin
title_full_unstemmed New Insights into Hemopexin-Binding to Hemin and Hemoglobin
title_short New Insights into Hemopexin-Binding to Hemin and Hemoglobin
title_sort new insights into hemopexin-binding to hemin and hemoglobin
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8998376/
https://www.ncbi.nlm.nih.gov/pubmed/35409149
http://dx.doi.org/10.3390/ijms23073789
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