title: Miniaturized Digestion and Extraction of Surface Proteins fromCandida albicansfollowing Treatment with Histatin 5 for Mass Spectrometry Analysis.
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 atμL volumes. Specifically,Candida albicanscells 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.
authors.affiliation: Department of Chemistry, Faculty of Science, University of Western Ontario, London, ON, Canada N6A 5B7.
authors.forename: Shirley
authors.initials: S
authors.lastname: Fan
authors.affiliation: Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1; Schulich Dentistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1.
authors.forename: Eduardo B
authors.identifier: 0000-0002-0372-6830
authors.initials: EB
authors.lastname: Moffa
authors.affiliation: Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1; Schulich Dentistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1.
authors.forename: Yizhi
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1; Schulich Dentistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1.
authors.forename: Walter L
authors.initials: WL
authors.lastname: Siqueira
authors.affiliation: Department of Chemistry, Faculty of Science, University of Western Ontario, London, ON, Canada N6A 5B7; Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada N6A 5C1.
authors.forename: Ken K-C
authors.identifier: 0000-0001-6933-2941
authors.initials: KK
authors.lastname: Yeung