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A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)

The Advanced Glycation and Lipoxidation End products (AGEs and ALEs) are a heterogeneous class of compounds derived from the non-enzymatic glycation or protein adduction by lipoxidation break-down products. The receptor for AGEs (RAGE) is involved in the progression of chronic diseases based on pers...

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Autores principales: Degani, Genny, Altomare, Alessandra A., Colzani, Mara, Martino, Caterina, Mazzolari, Angelica, Fritz, Guenter, Vistoli, Giulio, Popolo, Laura, Aldini, Giancarlo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5198869/
https://www.ncbi.nlm.nih.gov/pubmed/28013188
http://dx.doi.org/10.1016/j.redox.2016.12.017
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author Degani, Genny
Altomare, Alessandra A.
Colzani, Mara
Martino, Caterina
Mazzolari, Angelica
Fritz, Guenter
Vistoli, Giulio
Popolo, Laura
Aldini, Giancarlo
author_facet Degani, Genny
Altomare, Alessandra A.
Colzani, Mara
Martino, Caterina
Mazzolari, Angelica
Fritz, Guenter
Vistoli, Giulio
Popolo, Laura
Aldini, Giancarlo
author_sort Degani, Genny
collection PubMed
description The Advanced Glycation and Lipoxidation End products (AGEs and ALEs) are a heterogeneous class of compounds derived from the non-enzymatic glycation or protein adduction by lipoxidation break-down products. The receptor for AGEs (RAGE) is involved in the progression of chronic diseases based on persistent inflammatory state and oxidative stress. RAGE is a pattern recognition receptor (PRR) and the inhibition of the interaction with its ligands or of the ligand accumulation have a potential therapeutic effect. The N-terminal domain of RAGE, the V domain, is the major site of AGEs binding and is stabilized by the adjacent C1 domain. In this study, we set up an affinity assay relying on the extremely specific biological interaction AGEs ligands have for the VC1 domain. A glycosylated form of VC1, produced in the yeast Pichia pastoris, was attached to magnetic beads and used as insoluble affinity matrix (VC1-resin). The VC1 interaction assay was employed to isolate specific VC1 binding partners from in vitro generated AGE-albumins and modifications were identified/localized by mass spectrometry analysis. Interestingly, this method also led to the isolation of ALEs produced by malondialdehyde treatment of albumins. Computational studies provided a rational-based interpretation of the contacts established by specific modified residues and amino acids of the V domain. The validation of VC1-resin in capturing AGE-albumins from complex biological mixtures such as plasma and milk, may lead to the identification of new RAGE ligands potentially involved in pro-inflammatory and pro-fibrotic responses, independently of their structures or physical properties, and without the use of any covalent derivatization process. In addition, the method can be applied to the identification of antagonists of RAGE-ligand interaction.
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spelling pubmed-51988692017-01-04 A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE) Degani, Genny Altomare, Alessandra A. Colzani, Mara Martino, Caterina Mazzolari, Angelica Fritz, Guenter Vistoli, Giulio Popolo, Laura Aldini, Giancarlo Redox Biol Research Paper The Advanced Glycation and Lipoxidation End products (AGEs and ALEs) are a heterogeneous class of compounds derived from the non-enzymatic glycation or protein adduction by lipoxidation break-down products. The receptor for AGEs (RAGE) is involved in the progression of chronic diseases based on persistent inflammatory state and oxidative stress. RAGE is a pattern recognition receptor (PRR) and the inhibition of the interaction with its ligands or of the ligand accumulation have a potential therapeutic effect. The N-terminal domain of RAGE, the V domain, is the major site of AGEs binding and is stabilized by the adjacent C1 domain. In this study, we set up an affinity assay relying on the extremely specific biological interaction AGEs ligands have for the VC1 domain. A glycosylated form of VC1, produced in the yeast Pichia pastoris, was attached to magnetic beads and used as insoluble affinity matrix (VC1-resin). The VC1 interaction assay was employed to isolate specific VC1 binding partners from in vitro generated AGE-albumins and modifications were identified/localized by mass spectrometry analysis. Interestingly, this method also led to the isolation of ALEs produced by malondialdehyde treatment of albumins. Computational studies provided a rational-based interpretation of the contacts established by specific modified residues and amino acids of the V domain. The validation of VC1-resin in capturing AGE-albumins from complex biological mixtures such as plasma and milk, may lead to the identification of new RAGE ligands potentially involved in pro-inflammatory and pro-fibrotic responses, independently of their structures or physical properties, and without the use of any covalent derivatization process. In addition, the method can be applied to the identification of antagonists of RAGE-ligand interaction. Elsevier 2016-12-18 /pmc/articles/PMC5198869/ /pubmed/28013188 http://dx.doi.org/10.1016/j.redox.2016.12.017 Text en © 2016 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Research Paper
Degani, Genny
Altomare, Alessandra A.
Colzani, Mara
Martino, Caterina
Mazzolari, Angelica
Fritz, Guenter
Vistoli, Giulio
Popolo, Laura
Aldini, Giancarlo
A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)
title A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)
title_full A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)
title_fullStr A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)
title_full_unstemmed A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)
title_short A capture method based on the VC1 domain reveals new binding properties of the human receptor for advanced glycation end products (RAGE)
title_sort capture method based on the vc1 domain reveals new binding properties of the human receptor for advanced glycation end products (rage)
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5198869/
https://www.ncbi.nlm.nih.gov/pubmed/28013188
http://dx.doi.org/10.1016/j.redox.2016.12.017
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