Purpurin-induced changes in the proteome of Candida albicans
Antimicrobial agents/inhibitors, Fungi, Metabolism, Microbiology, Proteomics
Objectives: To identify the changes in protein abundance in Candida albicans after exposure to purpurin.
Methods: The optimal inhibitory dose of purpurin for proteomic analysis of C. albicans was determined by measuring fungal growth in YPD broth in the presence of a range of purpurin concentration (1-5 μg/mL) for 16 h. For proteomic analysis, total soluble proteins of purpurin-treated and untreated (DMSO only) fungal cells were extracted by mechanical disruption. In the first dimension IEF analysis, protein extract (200 μg) was focused on IPG strips (pH 3-10). The second dimension SDS-PAGE was performed on a 12% gel. The relative protein abundance was determined after silver staining. Highly reproducible spots showing (> 1.5-fold) up- or down-regulation were selected for identification using a MALDI-TOF mass spectrometer. The peak lists were searched against NCBI database using an in-house MASCOT searching engine.
Results: We identified 12 differentially (five up-regulated; seven down-regulated) expressed protein spots in the purpurin-treated C. albicans. These proteins are involved in stress and heat shock responses, TCA cycle, amino acid and aldehyde metabolism, and mitochondrial functions. Of special interest was a substantial increase (> 3 fold) of the cellular level of aryl alcohol dehydrogenase.
Conclusions: Purpurin induces changes in the proteome of C. albicans. Comparison of the differential protein expression patterns of the purpurin-treated C. albicans provides a better understanding of the antifungal mechanisms. The inhibitory effect of purpurin on Candida morphogenesis may be attributed to the up-regulation of aryl alcohol dehydrogenase, a crucial enzyme involved in the synthesis of quorum sensing molecules.
The 26th Annual Scientific Meeting of the International Association for Dental Research South-East Asia Division (IADR-SEA) and 23rd Annual Meeting of SEAADE, 2012 Oct 31 - Nov 4, Hong Kong
Tsang, W.,Bandara, H.,Samaranayake, L.,Tsang, P.,& Fong, W. (2012). Purpurin-induced changes in the proteome of Candida albicans. The 26th Annual Scientific Meeting of the International Association for Dental Research South-East Asia Division (IADR-SEA) and 23rd Annual Meeting of SEAADE, 2012 Oct 31 - Nov 4, Hong Kong. Retrieved from http://repository.vtc.edu.hk/thei-fac-gen-ed-sp/135