Table of Contents
International Journal of Proteomics
Volume 2016, Article ID 9812829, 12 pages
http://dx.doi.org/10.1155/2016/9812829
Research Article

Miniaturized Digestion and Extraction of Surface Proteins from Candida albicans following Treatment with Histatin 5 for Mass Spectrometry Analysis

1Department of Chemistry, Faculty of Science, University of Western Ontario, London, ON, Canada N6A 5B7
2Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1
3Schulich Dentistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1

Received 22 June 2016; Revised 25 October 2016; Accepted 31 October 2016

Academic Editor: Christian Huck

Copyright © 2016 Shirley Fan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

A common approach to isolate surface proteins from fungal and bacterial cells is to perform a proteolytic cleavage of proteins on the surface of intact cells suspended in solution. This paper describes miniaturization of this technique, in which cells are adhered on glass surfaces, and all sample treatments are conducted at μL volumes. Specifically, Candida albicans cells were attached onto HSA-coated glass slides. By depositing the appropriate reagent solutions on the adhered cells, we successfully performed cell washing, treatment with antifugal peptide, Histatin 5, and a proteolysis on intact cells with trypsin. The resulting peptides were subsequently analysed by mass spectrometry. In general, the data obtained was similar to that collected with suspended cells in much larger sample volumes. However, our miniaturized workflow offers the benefit of greatly reducing the consumption of cells and reagents.