An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins

Arsenic-binding proteins are under continuous research. Their identification and the elucidation of arsenic/ protein interaction mechanisms are important because the biological effects of these complexes may be related not only to arsenic but also to the arsenic/protein structure. Although many prot...

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Autores principales:	Femia, A.L., Temprana, C.F., Santos, J., Carbajal, M.L., Amor, M.S., Grasselli, M., Del Alonso, S.V.
Formato:	JOUR
Materias:	Arsenic carcinogenesis Circular dichroism Conformational studies Drug delivery systems Thioredoxin 4 aminophenylarsine oxide arsenic arsenic binding protein binding protein fluorescein isothiocyanate fluorescent dye thioredoxin 1 unclassified drug article atomic absorption circular dichroism enzyme activity fluorescence polyacrylamide gel electrophoresis protein folding protein interaction protein structure Arsenic Arsenicals Carrier Proteins Fluorescein-5-isothiocyanate Fluorescent Dyes Kinetics Models, Molecular Protein Binding Protein Conformation Protein Unfolding Spectrum Analysis Temperature Thioredoxins
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_15723887_v31_n8_p656_Femia
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_15723887_v31_n8_p656_Femia
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spelling	todo:paper_15723887_v31_n8_p656_Femia2023-10-03T16:27:20Z An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins Femia, A.L. Temprana, C.F. Santos, J. Carbajal, M.L. Amor, M.S. Grasselli, M. Del Alonso, S.V. Arsenic carcinogenesis Circular dichroism Conformational studies Drug delivery systems Thioredoxin 4 aminophenylarsine oxide arsenic arsenic binding protein binding protein fluorescein isothiocyanate fluorescent dye thioredoxin 1 unclassified drug article atomic absorption circular dichroism enzyme activity fluorescence polyacrylamide gel electrophoresis protein folding protein interaction protein structure Arsenic Arsenicals Carrier Proteins Fluorescein-5-isothiocyanate Fluorescent Dyes Kinetics Models, Molecular Protein Binding Protein Conformation Protein Unfolding Spectrum Analysis Temperature Thioredoxins Arsenic-binding proteins are under continuous research. Their identification and the elucidation of arsenic/ protein interaction mechanisms are important because the biological effects of these complexes may be related not only to arsenic but also to the arsenic/protein structure. Although many proteins bearing a CXXC motif have been found to bind arsenic in vivo, new tools are necessary to identify new arsenic targets and allow research on protein/ arsenic complexes. In this work, we analyzed the performance of the fluorescent compound APAO-FITC (synthesized from p-aminophenylarsenoxide, APAO, and fluorescein isothiocyanate, FITC) in arsenic/protein binding assays using thioredoxin 1 (Trx) as an arsenic-binding protein model. The Trx-APAO-FITC complex was studied through different spectroscopic techniques involving UV-Vis, fluorescence, atomic absorption, infrared and circular dichroism. Our results show that APAO-FITC binds efficiently and specifically to the Trx binding site, labeling the protein fluorescently, without altering its structure and activity. In summary, we were able to study a protein/ arsenic complex model, using APAO-FITC as a labeling probe. The use of APAO-FITC in the identification of different protein and cell targets, as well as in in vivo biodistribution studies, conformational studies of arsenicbinding proteins, and studies for the design of drug delivery systems for arsenic anti-cancer therapies, is highly promising. © Springer Science+Business Media, LLC 2012. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_15723887_v31_n8_p656_Femia
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Arsenic carcinogenesis Circular dichroism Conformational studies Drug delivery systems Thioredoxin 4 aminophenylarsine oxide arsenic arsenic binding protein binding protein fluorescein isothiocyanate fluorescent dye thioredoxin 1 unclassified drug article atomic absorption circular dichroism enzyme activity fluorescence polyacrylamide gel electrophoresis protein folding protein interaction protein structure Arsenic Arsenicals Carrier Proteins Fluorescein-5-isothiocyanate Fluorescent Dyes Kinetics Models, Molecular Protein Binding Protein Conformation Protein Unfolding Spectrum Analysis Temperature Thioredoxins
spellingShingle	Arsenic carcinogenesis Circular dichroism Conformational studies Drug delivery systems Thioredoxin 4 aminophenylarsine oxide arsenic arsenic binding protein binding protein fluorescein isothiocyanate fluorescent dye thioredoxin 1 unclassified drug article atomic absorption circular dichroism enzyme activity fluorescence polyacrylamide gel electrophoresis protein folding protein interaction protein structure Arsenic Arsenicals Carrier Proteins Fluorescein-5-isothiocyanate Fluorescent Dyes Kinetics Models, Molecular Protein Binding Protein Conformation Protein Unfolding Spectrum Analysis Temperature Thioredoxins Femia, A.L. Temprana, C.F. Santos, J. Carbajal, M.L. Amor, M.S. Grasselli, M. Del Alonso, S.V. An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
topic_facet	Arsenic carcinogenesis Circular dichroism Conformational studies Drug delivery systems Thioredoxin 4 aminophenylarsine oxide arsenic arsenic binding protein binding protein fluorescein isothiocyanate fluorescent dye thioredoxin 1 unclassified drug article atomic absorption circular dichroism enzyme activity fluorescence polyacrylamide gel electrophoresis protein folding protein interaction protein structure Arsenic Arsenicals Carrier Proteins Fluorescein-5-isothiocyanate Fluorescent Dyes Kinetics Models, Molecular Protein Binding Protein Conformation Protein Unfolding Spectrum Analysis Temperature Thioredoxins
description	Arsenic-binding proteins are under continuous research. Their identification and the elucidation of arsenic/ protein interaction mechanisms are important because the biological effects of these complexes may be related not only to arsenic but also to the arsenic/protein structure. Although many proteins bearing a CXXC motif have been found to bind arsenic in vivo, new tools are necessary to identify new arsenic targets and allow research on protein/ arsenic complexes. In this work, we analyzed the performance of the fluorescent compound APAO-FITC (synthesized from p-aminophenylarsenoxide, APAO, and fluorescein isothiocyanate, FITC) in arsenic/protein binding assays using thioredoxin 1 (Trx) as an arsenic-binding protein model. The Trx-APAO-FITC complex was studied through different spectroscopic techniques involving UV-Vis, fluorescence, atomic absorption, infrared and circular dichroism. Our results show that APAO-FITC binds efficiently and specifically to the Trx binding site, labeling the protein fluorescently, without altering its structure and activity. In summary, we were able to study a protein/ arsenic complex model, using APAO-FITC as a labeling probe. The use of APAO-FITC in the identification of different protein and cell targets, as well as in in vivo biodistribution studies, conformational studies of arsenicbinding proteins, and studies for the design of drug delivery systems for arsenic anti-cancer therapies, is highly promising. © Springer Science+Business Media, LLC 2012.
format	JOUR
author	Femia, A.L. Temprana, C.F. Santos, J. Carbajal, M.L. Amor, M.S. Grasselli, M. Del Alonso, S.V.
author_facet	Femia, A.L. Temprana, C.F. Santos, J. Carbajal, M.L. Amor, M.S. Grasselli, M. Del Alonso, S.V.
author_sort	Femia, A.L.
title	An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
title_short	An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
title_full	An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
title_fullStr	An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
title_full_unstemmed	An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
title_sort	arsenic fluorescent compound as a novel probe to study arsenic-binding proteins
url	http://hdl.handle.net/20.500.12110/paper_15723887_v31_n8_p656_Femia
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An arsenic fluorescent compound as a novel probe to study arsenic-binding proteins

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