Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection
Capillary electrophoresis (CE) coupled with laser-induced fluorescence detection (LIF) has allowed to obtain protein fingerprints, which have demonstrated to be useful in microorganisms characterization. In this work, protein fingerprints of two species of Staphylococcus grown in different culture media and submitted to temperature and nitrosative stress were studied by CE-LIF. After the growth of the bacteria, protein extracts were obtained by cell lysis using sonication. The water-soluble fraction of these lysates was derivatized on-capillary with a fluorogenic dye, 3-(2-furoyl)quinoline-2-carboxaldehyde. The fluorescent products were analyzed by CE using phosphate buffer containing submicellar concentrations of sodium pentanesulfate and detected by LIF. Different protein fingerprints were obtained depending on the bacterial specie studied, indicating the usefulness of this method for the identification of different species of the same bacterial genus. It was also demonstrated that the CE protein fingerprints were dependent on the culture conditions, such as growth medium, or on stressing conditions, such as heat shock or nitrosative stress
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Language: | English |
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Elsevier
2010-01-18
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Subjects: | Capillary electrophoresis, Laser-induced fluorescence detection, Protein fingerprinting;, Staphylococcus, |
Online Access: | http://hdl.handle.net/10261/41043 http://dx.doi.org/10.13039/501100004837 http://dx.doi.org/10.13039/100012818 |
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dig-icvv-es-10261-410432017-03-07T16:17:12Z Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection Veledo, María Teresa Pelaez-Lorenzo, Cristina González García, Ramón Frutos, Mercedes de Díez-Masa, José Carlos Ministerio de Ciencia e Innovación (España) Comunidad de Madrid Fundación Domingo Martínez Capillary electrophoresis Laser-induced fluorescence detection Protein fingerprinting; Staphylococcus Capillary electrophoresis (CE) coupled with laser-induced fluorescence detection (LIF) has allowed to obtain protein fingerprints, which have demonstrated to be useful in microorganisms characterization. In this work, protein fingerprints of two species of Staphylococcus grown in different culture media and submitted to temperature and nitrosative stress were studied by CE-LIF. After the growth of the bacteria, protein extracts were obtained by cell lysis using sonication. The water-soluble fraction of these lysates was derivatized on-capillary with a fluorogenic dye, 3-(2-furoyl)quinoline-2-carboxaldehyde. The fluorescent products were analyzed by CE using phosphate buffer containing submicellar concentrations of sodium pentanesulfate and detected by LIF. Different protein fingerprints were obtained depending on the bacterial specie studied, indicating the usefulness of this method for the identification of different species of the same bacterial genus. It was also demonstrated that the CE protein fingerprints were dependent on the culture conditions, such as growth medium, or on stressing conditions, such as heat shock or nitrosative stress Our acknowledgement to Spanish Ministry of Science and Innovation (PATSENS project, #PSE-01000-2008-6), to Comunidad de Madrid (S-GEN/0247/2006 project) and to Domingo Martínez Foundation for financial support Peer reviewed 2011-10-13T10:28:38Z 2011-10-13T10:28:38Z 2010-01-18 artículo http://purl.org/coar/resource_type/c_6501 Analytica Chimica Acta 68(1): 81-86 (2010) 0003-2670 http://hdl.handle.net/10261/41043 10.1016/j.aca.2009.10.053 http://dx.doi.org/10.13039/501100004837 http://dx.doi.org/10.13039/100012818 en http://dx.doi.org/10.1016/j.aca.2009.10.053 none Elsevier |
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Capillary electrophoresis Laser-induced fluorescence detection Protein fingerprinting; Staphylococcus Capillary electrophoresis Laser-induced fluorescence detection Protein fingerprinting; Staphylococcus |
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Capillary electrophoresis Laser-induced fluorescence detection Protein fingerprinting; Staphylococcus Capillary electrophoresis Laser-induced fluorescence detection Protein fingerprinting; Staphylococcus Veledo, María Teresa Pelaez-Lorenzo, Cristina González García, Ramón Frutos, Mercedes de Díez-Masa, José Carlos Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
description |
Capillary electrophoresis (CE) coupled with laser-induced fluorescence detection (LIF) has allowed to obtain protein fingerprints, which have demonstrated to be useful in microorganisms characterization. In this work, protein fingerprints of two species of Staphylococcus grown in different culture media and submitted to temperature and nitrosative stress were studied by CE-LIF. After the growth of the bacteria, protein extracts were obtained by cell lysis using sonication. The water-soluble fraction of these lysates was derivatized on-capillary with a fluorogenic dye, 3-(2-furoyl)quinoline-2-carboxaldehyde. The fluorescent products were analyzed by CE using phosphate buffer containing submicellar concentrations of sodium pentanesulfate and detected by LIF. Different protein fingerprints were obtained depending on the bacterial specie studied, indicating the usefulness of this method for the identification of different species of the same bacterial genus. It was also demonstrated that the CE protein fingerprints were dependent on the culture conditions, such as growth medium, or on stressing conditions, such as heat shock or nitrosative stress |
author2 |
Ministerio de Ciencia e Innovación (España) |
author_facet |
Ministerio de Ciencia e Innovación (España) Veledo, María Teresa Pelaez-Lorenzo, Cristina González García, Ramón Frutos, Mercedes de Díez-Masa, José Carlos |
format |
artículo |
topic_facet |
Capillary electrophoresis Laser-induced fluorescence detection Protein fingerprinting; Staphylococcus |
author |
Veledo, María Teresa Pelaez-Lorenzo, Cristina González García, Ramón Frutos, Mercedes de Díez-Masa, José Carlos |
author_sort |
Veledo, María Teresa |
title |
Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
title_short |
Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
title_full |
Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
title_fullStr |
Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
title_full_unstemmed |
Protein fingerprinting of Staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
title_sort |
protein fingerprinting of staphylococcus species by capillary electrophoresis with on-capillary derivatization and laser-induced fluorescence detection |
publisher |
Elsevier |
publishDate |
2010-01-18 |
url |
http://hdl.handle.net/10261/41043 http://dx.doi.org/10.13039/501100004837 http://dx.doi.org/10.13039/100012818 |
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