Cargando…
FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts
p53 over expression in yeast results in cell death with typical markers of apoptosis such as DNA fragmentation and phosphatidylserine externalization. We aimed to substitute/supplement classical fluorescent techniques (TUNEL, Annexin V, ROS detection) usually used to detect biochemical changes occur...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507515/ https://www.ncbi.nlm.nih.gov/pubmed/28704406 http://dx.doi.org/10.1371/journal.pone.0180680 |
_version_ | 1783249750859972608 |
---|---|
author | Mihoubi, Wafa Sahli, Emna Gargouri, Ali Amiel, Caroline |
author_facet | Mihoubi, Wafa Sahli, Emna Gargouri, Ali Amiel, Caroline |
author_sort | Mihoubi, Wafa |
collection | PubMed |
description | p53 over expression in yeast results in cell death with typical markers of apoptosis such as DNA fragmentation and phosphatidylserine externalization. We aimed to substitute/supplement classical fluorescent techniques (TUNEL, Annexin V, ROS detection) usually used to detect biochemical changes occurring during yeast apoptosis mediated by p53 over expression and the effect of anti-apoptotic purified molecules from Nigel (Nigella sativa) extracts on these same yeasts by the label free technique of FTIR spectroscopy. The comparison of the entire IR spectra highlighted clear modifications between apoptotic p53-expressing yeasts and normal ones. More precisely, DNA damage was detected by the decrease of band intensities at 1079 and 1048 cm(-1). While phosphatidylserine exposure was followed by the increase of νsCH(2) and νasCH(2) bands of unsaturated fatty acids that were exhibited at 2855 and 2926 cm(-1), and the appearance of the C = O ester functional group band at 1740 cm(-1). In a second step, this FTIR approach was used to estimate the effect of a purified fraction of the Nigel extract. The modulation of band intensities specific to DNA and membrane status was in agreement with apoptosis supression in presence of the Nigel extracts. FTIR spectroscopy is thus proven to be a very reliable technique to monitor the apoptotic cell death in yeast and to be used as a means of evaluating the biomolecules effect on yeast survival. |
format | Online Article Text |
id | pubmed-5507515 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-55075152017-07-25 FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts Mihoubi, Wafa Sahli, Emna Gargouri, Ali Amiel, Caroline PLoS One Research Article p53 over expression in yeast results in cell death with typical markers of apoptosis such as DNA fragmentation and phosphatidylserine externalization. We aimed to substitute/supplement classical fluorescent techniques (TUNEL, Annexin V, ROS detection) usually used to detect biochemical changes occurring during yeast apoptosis mediated by p53 over expression and the effect of anti-apoptotic purified molecules from Nigel (Nigella sativa) extracts on these same yeasts by the label free technique of FTIR spectroscopy. The comparison of the entire IR spectra highlighted clear modifications between apoptotic p53-expressing yeasts and normal ones. More precisely, DNA damage was detected by the decrease of band intensities at 1079 and 1048 cm(-1). While phosphatidylserine exposure was followed by the increase of νsCH(2) and νasCH(2) bands of unsaturated fatty acids that were exhibited at 2855 and 2926 cm(-1), and the appearance of the C = O ester functional group band at 1740 cm(-1). In a second step, this FTIR approach was used to estimate the effect of a purified fraction of the Nigel extract. The modulation of band intensities specific to DNA and membrane status was in agreement with apoptosis supression in presence of the Nigel extracts. FTIR spectroscopy is thus proven to be a very reliable technique to monitor the apoptotic cell death in yeast and to be used as a means of evaluating the biomolecules effect on yeast survival. Public Library of Science 2017-07-12 /pmc/articles/PMC5507515/ /pubmed/28704406 http://dx.doi.org/10.1371/journal.pone.0180680 Text en © 2017 Mihoubi et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Mihoubi, Wafa Sahli, Emna Gargouri, Ali Amiel, Caroline FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts |
title | FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts |
title_full | FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts |
title_fullStr | FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts |
title_full_unstemmed | FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts |
title_short | FTIR spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by Nigella sativa extracts |
title_sort | ftir spectroscopy of whole cells for the monitoring of yeast apoptosis mediated by p53 over-expression and its suppression by nigella sativa extracts |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507515/ https://www.ncbi.nlm.nih.gov/pubmed/28704406 http://dx.doi.org/10.1371/journal.pone.0180680 |
work_keys_str_mv | AT mihoubiwafa ftirspectroscopyofwholecellsforthemonitoringofyeastapoptosismediatedbyp53overexpressionanditssuppressionbynigellasativaextracts AT sahliemna ftirspectroscopyofwholecellsforthemonitoringofyeastapoptosismediatedbyp53overexpressionanditssuppressionbynigellasativaextracts AT gargouriali ftirspectroscopyofwholecellsforthemonitoringofyeastapoptosismediatedbyp53overexpressionanditssuppressionbynigellasativaextracts AT amielcaroline ftirspectroscopyofwholecellsforthemonitoringofyeastapoptosismediatedbyp53overexpressionanditssuppressionbynigellasativaextracts |