title: Tracing putative trafficking of the glycolytic enzyme enolase via SNARE-driven unconventional secretion.
abstract: Glycolytic enzymes are cytosolic proteins, but they also play important extracellular roles in cell-cell communication and infection. We used Saccharomyces cerevisiae to analyze the secretory pathway of some of these enzymes, including enolase, phosphoglucose isomerase, triose phosphate isomerase, and fructose 1,6-bisphosphate aldolase. Enolase, phosphoglucose isomerase, and an N-terminal 28-amino-acid-long fragment of enolase were secreted in a sec23-independent manner. The enhanced green fluorescent protein (EGFP)-conjugated enolase fragment formed cellular foci, some of which were found at the cell periphery. Therefore, we speculated that an overview of the secretory pathway could be gained by investigating the colocalization of the enolase fragment with intracellular proteins. The DsRed-conjugated enolase fragment colocalized with membrane proteins at the cis-Golgi complex, nucleus, endosome, and plasma membrane, but not the mitochondria. In addition, the secretion of full-length enolase was inhibited in a knockout mutant of the intracellular SNARE protein-coding gene TLG2. Our results suggest that enolase is secreted via a SNARE-dependent secretory pathway in S. cerevisiae.
authors: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto, Japan.
authors: Natsuko
authors: N
authors: Miura
authors: Aya
authors: A
authors: Kirino
authors: Satoshi
authors: S
authors: Endo
authors: Hironobu
authors: H
authors: Morisaka
authors: Kouichi
authors: K
authors: Kuroda
authors: Masahiro
authors: M
authors: Takagi
authors: Mitsuyoshi
authors: M
authors: Ueda
mesh_terms: D002462:Cell Membrane / Q000737:chemistry; D011992:Endosomes / Q000737:chemistry; D017930:Genes, Reporter / Q000235:genetics; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism; D006056:Golgi Apparatus / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008928:Mitochondria / Q000737:chemistry; D009154:Mutation; D010751:Phosphopyruvate Hydratase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D050765:Qa-SNARE Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D055571:Secretory Pathway
chemical_list: D008164:Luminescent Proteins; D050765:Qa-SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; C112252:TLG2 protein, S cerevisiae; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins; D010751:Phosphopyruvate Hydratase; C516797:enolase 2, S cerevisiae; D005956:Glucose-6-Phosphate Isomerase