title: Identification of Secreted Candida Proteins Using Mass Spectrometry.
abstract: Analysis of fungal secretomes using mass spectrometry is a useful technique in cell biology. Knowledge of the secretome of a human fungal pathogen may yield important information of host-pathogen interactions and may be useful for identifying vaccines candidates or diagnostic markers for antifungal strategies. In this chapter, with a main focus on sample preparation aspects, we describe the methodology that we apply for gel-independent batch identification and quantification of proteins that are secreted during growth in liquid cultures. Using these techniques with Candida and other yeast species, the majority of the identified proteins are classical secretory proteins and cell wall proteins containing N-terminal signal peptides for secretion, although dependent on sample preparation quality and the mass spectrometric analysis also usually, a number of nonsecretory proteins are identified.
authors.affiliation: Regional Center for Biomedical Research, Albacete Science & Technology Park, University of Castilla-La Mancha, Calle Almansa 14, 02008, Albacete, Spain.
authors.forename: Emilia
authors.initials: E
authors.lastname: Gómez-Molero
authors.affiliation: Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH, Amsterdam, The Netherlands.
authors.forename: Henk L
authors.initials: HL
authors.lastname: Dekker
authors.affiliation: Regional Center for Biomedical Research, Albacete Science & Technology Park, University of Castilla-La Mancha, Calle Almansa 14, 02008, Albacete, Spain.
authors.forename: Albert D
authors.initials: AD
authors.lastname: de Boer
authors.affiliation: Regional Center for Biomedical Research, Albacete Science & Technology Park, University of Castilla-La Mancha, Calle Almansa 14, 02008, Albacete, Spain. piet.degroot@uclm.es.
authors.forename: Piet W J
authors.initials: PW
authors.lastname: de Groot
mesh_terms: D002175:Candida / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D020543:Proteome; D040901:Proteomics / Q000379:methods
chemical_list: D005656:Fungal Proteins; D020543:Proteome
keywords: Candida; LC/MS/MS; Mass spectrometry; Secretome; Signal peptide; Trypsin