Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte

Zein proteins are a complex mixture of polypetides that belong to the alcohol soluble storage proteins group (prolamines) in corn. These proteins constitute about 50-60% of the total endosperm protein and are classified in different groups on the basis of differences in their solubility and sequence...

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Authors: Erny, Guillaume L., Marina, María Luisa, Cifuentes, Alejandro
Format: article
Publication Date:2007
Country:España
Institution:Consejo Superior de Investigaciones Científicas (CSIC)
Repository:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/12921
Online Access:http://hdl.handle.net/10261/12921
Access Level:Open access
Keyword:Coatings
protein separation
ammonium gradient
Corn
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spelling Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyteErny, Guillaume L.Marina, María LuisaCifuentes, AlejandroCoatingsprotein separationammonium gradientCornZein proteins are a complex mixture of polypetides that belong to the alcohol soluble storage proteins group (prolamines) in corn. These proteins constitute about 50-60% of the total endosperm protein and are classified in different groups on the basis of differences in their solubility and sequence. Among them, zein proteins are considered the majority group showing a high tendency to aggregate what makes very difficult their analysis by any analytical method. Thus, capillary zone electrophoresis (CZE) of these proteins require the use of very complex background electrolytes (BGEs) non-compatible with on-line ESI-MS analysis. The aim of this work was to find a new BGE for the CZE separation of zein protein fully compatible with ESI-MS while providing further light on the complex CZE separation of aggregatable proteins. Thus, it is demonstrated in this work that efficient and reproducible CZE separations of zein proteins can be achieved by using a BGE composed of water, acetonitrile, formic acid and ammonium hydroxide. Besides, it is shown that zeins analysis is significantly improved by including the effect of an ammonium gradient during their separation. It is experimentally verified that the ammonium gradient can easily be achieved in CZE by either working with a sample zone with a low concentration of ammonium and a background electrolyte (BGE) with a high concentration, or conversely, working with a sample zone with high ammonium concentration and a BGE with low concentration of ammonium, giving rise in both cases to a significant improvement in the CZE separation of these proteins. It is demonstrated that this procedure can give rise to efficiency improvements of up to 20-folds in the CZE separation of zein proteins. Under optimized conditions, 20 proteins could be separated with average efficiencies higher than 400000 theoretical plates/m. Some possible explanations of this effect are discussed including stacking, protein-capillary wall adsorption, protein solubility and protein-salt interactions.Authors are grateful to the AGL2005-05320-C02-01 Project (Ministerio de Educacion y Ciencia) and the S-505/AGR-0153 Project (Comunidad Autonoma de Madrid) for financial support of this work.Peer reviewedWiley-VCH200920092007info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501331776 bytesapplication/mswordhttp://hdl.handle.net/10261/12921reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés10.1002/elps.200790062info:eu-repo/semantics/openAccessoai:digital.csic.es:10261/129212026-05-22T06:33:51Z
dc.title.none.fl_str_mv Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
title Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
spellingShingle Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
Erny, Guillaume L.
Coatings
protein separation
ammonium gradient
Corn
title_short Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
title_full Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
title_fullStr Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
title_full_unstemmed Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
title_sort Reproducible and efficient separation of aggregatable zein proteins by capillary zone electrophoresis using a volatile background electrolyte
dc.creator.none.fl_str_mv Erny, Guillaume L.
Marina, María Luisa
Cifuentes, Alejandro
author Erny, Guillaume L.
author_facet Erny, Guillaume L.
Marina, María Luisa
Cifuentes, Alejandro
author_role author
author2 Marina, María Luisa
Cifuentes, Alejandro
author2_role author
author
dc.subject.none.fl_str_mv Coatings
protein separation
ammonium gradient
Corn
topic Coatings
protein separation
ammonium gradient
Corn
description Zein proteins are a complex mixture of polypetides that belong to the alcohol soluble storage proteins group (prolamines) in corn. These proteins constitute about 50-60% of the total endosperm protein and are classified in different groups on the basis of differences in their solubility and sequence. Among them, zein proteins are considered the majority group showing a high tendency to aggregate what makes very difficult their analysis by any analytical method. Thus, capillary zone electrophoresis (CZE) of these proteins require the use of very complex background electrolytes (BGEs) non-compatible with on-line ESI-MS analysis. The aim of this work was to find a new BGE for the CZE separation of zein protein fully compatible with ESI-MS while providing further light on the complex CZE separation of aggregatable proteins. Thus, it is demonstrated in this work that efficient and reproducible CZE separations of zein proteins can be achieved by using a BGE composed of water, acetonitrile, formic acid and ammonium hydroxide. Besides, it is shown that zeins analysis is significantly improved by including the effect of an ammonium gradient during their separation. It is experimentally verified that the ammonium gradient can easily be achieved in CZE by either working with a sample zone with a low concentration of ammonium and a background electrolyte (BGE) with a high concentration, or conversely, working with a sample zone with high ammonium concentration and a BGE with low concentration of ammonium, giving rise in both cases to a significant improvement in the CZE separation of these proteins. It is demonstrated that this procedure can give rise to efficiency improvements of up to 20-folds in the CZE separation of zein proteins. Under optimized conditions, 20 proteins could be separated with average efficiencies higher than 400000 theoretical plates/m. Some possible explanations of this effect are discussed including stacking, protein-capillary wall adsorption, protein solubility and protein-salt interactions.
publishDate 2007
dc.date.none.fl_str_mv 2007
2009
2009
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/12921
url http://hdl.handle.net/10261/12921
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv 10.1002/elps.200790062
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 331776 bytes
application/msword
dc.publisher.none.fl_str_mv Wiley-VCH
publisher.none.fl_str_mv Wiley-VCH
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
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