Staff View: Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy

Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy

Candidiasis is a fungal infection leading to significant morbidity and mortality in immunocompromised patients. Aureobasidin (AbA), a cyclic depsipeptide antifungal drug, has been reported to be effective against Candida albicans but is yet to be used for therapy. Although AbA is presumed to targ...

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Main Author:	Komathy, Munusamy
Format:	Thesis
Published:	2019
Subjects:	R Medicine (General)
Online Access:	http://studentsrepo.um.edu.my/11455/4/komathy.pdf http://studentsrepo.um.edu.my/11455/
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spelling	my.um.stud.114552022-01-03T23:23:26Z Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy Komathy, Munusamy R Medicine (General) Candidiasis is a fungal infection leading to significant morbidity and mortality in immunocompromised patients. Aureobasidin (AbA), a cyclic depsipeptide antifungal drug, has been reported to be effective against Candida albicans but is yet to be used for therapy. Although AbA is presumed to target the fungal sphingolipid biosynthesis, the exact mechanism of action has yet to be defined. As a prerequisite to study the mechanism of action of this antifungal drug, this study investigated changes in the proteome of planktonic and biofilm cultures of C. albicans upon exposure to AbA. In this study, the whole cell extracts of biofilm and planktonic cultures of C. albicans strain SC5314 pre- and post-exposure to AbA were subjected to liquid chromatography mass spectrometry (LC-MS/MS) analysis. Using sequence homology search tool (SPIDER) of PEAKS software (ver. 7.5), proteins were identified based on the setting of at least one unique peptide matched and a significance (-10lgP) value of >20. The proteome prior to AbA exposure demonstrated a total of 203 and 533 proteins from the planktonic and biofilm cultures, respectively, with a false discovery rate of ≤1%. The biological process, molecular function, and subcellular localization of biofilm and planktonic proteins were annotated using Go Slim Mapper of the online Candida Genome Database. Classification of biological processes demonstrated greater differences in the number of biofilm proteins associated with the regulation of biological processes (n=65), organelle organization (n=57), transport (n=55), stress response (n=47), filamentous growth (n=30), chemical response (n=30), lipid metabolism (n=23) and carbohydrate metabolism (n=20) compared to planktonic culture. As for molecular function, no major difference was observed in both cultures, iv with approximately 10-20% of the proteins are associated with hydrolase, transferase and protein binding activities. On subcellular localization, 68.7 and 66.4% proteins of planktonic and biofilm cultures were predicted to be localized in cytoplasm. A total of 293 and 374 proteins were annotated in the AbA-treated planktonic and biofilm cultures with sub-inhinbitory AbA concentration (1 µg/ml) for 2.5 hours. A lower percentage of proteins annotated for organelle organization in the AbA-treated planktonic culture (18.1%) as compared to the dimethyl sulfoxide (DMSO)-treated planktonic culture (23.2%). There was a 5% increase in the proteins annotated for translation in AbAtreated planktonic culture. AbA-treated biofilm culture has shown reduction in the percentages of proteins annotated for vesicle-mediated transport (from 8.4 to 5.3%) and filamentous growth (15.8 to 12.8%), and an increase in the percentage of proteins annotated for carbohydrate metabolic process (8.2 to 12.0%). The regulation of cellular amide metabolic process and actin cytoskeleton organization are two biological processes shared by AbA-treated planktonic and biofilm cultures. This study also reviewed some C. albicans proteins which are annotated for pathogenesis, biofilm formation, lipid metabolic process and filamentous growth in response to AbA. Further exploration of the drug-affected proteins may aid in the search for potential drug target in the treatment of candidiasis. 2019 Thesis NonPeerReviewed application/pdf http://studentsrepo.um.edu.my/11455/4/komathy.pdf Komathy, Munusamy (2019) Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy. Masters thesis, Universiti Malaya. http://studentsrepo.um.edu.my/11455/
institution	Universiti Malaya
building	UM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Malaya
content_source	UM Student Repository
url_provider	http://studentsrepo.um.edu.my/
topic	R Medicine (General)
spellingShingle	R Medicine (General) Komathy, Munusamy Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy
description	Candidiasis is a fungal infection leading to significant morbidity and mortality in immunocompromised patients. Aureobasidin (AbA), a cyclic depsipeptide antifungal drug, has been reported to be effective against Candida albicans but is yet to be used for therapy. Although AbA is presumed to target the fungal sphingolipid biosynthesis, the exact mechanism of action has yet to be defined. As a prerequisite to study the mechanism of action of this antifungal drug, this study investigated changes in the proteome of planktonic and biofilm cultures of C. albicans upon exposure to AbA. In this study, the whole cell extracts of biofilm and planktonic cultures of C. albicans strain SC5314 pre- and post-exposure to AbA were subjected to liquid chromatography mass spectrometry (LC-MS/MS) analysis. Using sequence homology search tool (SPIDER) of PEAKS software (ver. 7.5), proteins were identified based on the setting of at least one unique peptide matched and a significance (-10lgP) value of >20. The proteome prior to AbA exposure demonstrated a total of 203 and 533 proteins from the planktonic and biofilm cultures, respectively, with a false discovery rate of ≤1%. The biological process, molecular function, and subcellular localization of biofilm and planktonic proteins were annotated using Go Slim Mapper of the online Candida Genome Database. Classification of biological processes demonstrated greater differences in the number of biofilm proteins associated with the regulation of biological processes (n=65), organelle organization (n=57), transport (n=55), stress response (n=47), filamentous growth (n=30), chemical response (n=30), lipid metabolism (n=23) and carbohydrate metabolism (n=20) compared to planktonic culture. As for molecular function, no major difference was observed in both cultures, iv with approximately 10-20% of the proteins are associated with hydrolase, transferase and protein binding activities. On subcellular localization, 68.7 and 66.4% proteins of planktonic and biofilm cultures were predicted to be localized in cytoplasm. A total of 293 and 374 proteins were annotated in the AbA-treated planktonic and biofilm cultures with sub-inhinbitory AbA concentration (1 µg/ml) for 2.5 hours. A lower percentage of proteins annotated for organelle organization in the AbA-treated planktonic culture (18.1%) as compared to the dimethyl sulfoxide (DMSO)-treated planktonic culture (23.2%). There was a 5% increase in the proteins annotated for translation in AbAtreated planktonic culture. AbA-treated biofilm culture has shown reduction in the percentages of proteins annotated for vesicle-mediated transport (from 8.4 to 5.3%) and filamentous growth (15.8 to 12.8%), and an increase in the percentage of proteins annotated for carbohydrate metabolic process (8.2 to 12.0%). The regulation of cellular amide metabolic process and actin cytoskeleton organization are two biological processes shared by AbA-treated planktonic and biofilm cultures. This study also reviewed some C. albicans proteins which are annotated for pathogenesis, biofilm formation, lipid metabolic process and filamentous growth in response to AbA. Further exploration of the drug-affected proteins may aid in the search for potential drug target in the treatment of candidiasis.
format	Thesis
author	Komathy, Munusamy
author_facet	Komathy, Munusamy
author_sort	Komathy, Munusamy
title	Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy
title_short	Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy
title_full	Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy
title_fullStr	Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy
title_full_unstemmed	Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy
title_sort	protein profiling of candida albicans planktonic and biofilm cultures upon exposure to aureobasidin / komathy munusamy
publishDate	2019
url	http://studentsrepo.um.edu.my/11455/4/komathy.pdf http://studentsrepo.um.edu.my/11455/
_version_	1738506486589423616
score	13.211869

Protein profiling of Candida albicans planktonic and biofilm cultures upon exposure to Aureobasidin / Komathy Munusamy

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