title: Evaluation of unconventional protein secretion in Saccharomyces cerevisiae.
abstract: Recent development of large-scale analyses such as the secretome analysis has enabled the discovery of a vast number of intracellular proteins that are secreted outside the cell. Often, these proteins do not contain any known signal sequence required for conventional protein secretion. In order to avoid misidentification of such "leaked" proteins as "secreted" proteins, reconstructing the process of protein secretion is essential. Here, we describe methods for the detection of reconstructed unconventional protein secretion and determination of regulatory proteins of secretion in Saccharomyces cerevisiae. We show that conjugating target proteins with a tag-sequence and utilizing various reagents and tools can facilitate quantitative detection of the secretion of target proteins. We expect that these methods will reveal novel unconventional secretion pathways of proteins.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
authors.forename: Natsuko
authors.initials: N
authors.lastname: Miura
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
mesh_terms: D005110:Extracellular Space / Q000378:metabolism; D015870:Gene Expression; D055786:Gene Knockout Techniques; D042541:Intracellular Space / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D055571:Secretory Pathway; D014170:Transformation, Genetic
chemical_list: D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins