Protein transport in the permeabilized cell of Schizosaccharomyces pombe.
We reconstituted a protein translocation-transport system composed of permeabilized spheroplasts (P-cells) of the fission yeast Schizosaccharomyces pombe and the precursor of alpha sex pheromone, prepro-alpha-factor of the budding yeast Saccharomyces cerevisiae. We found that P-cells prepared from the spheroplasts formed in 0.7M KCl as an osmotic stabilizer had the activity to transport pro-alpha-factor to the Golgi apparatus. Electron microscopic observations showed that membranes were preserved more intact in the P-cells prepared from the spheroplasts formed in 0.7M KCl than in 0.7M sorbitol. A glycoprotein of S. pombe contains galactose residues, and we detected incorporation of radiolabeled galactose residues into the anti-prepro-alpha-factor immunoprecipitable fractions in this S. pombe system, but not in the S. cerevisiae system. This paper reports that a heterologous system of in vitro protein transport was performed, and prepro-alpha-factor has the signals necessary for early steps of the transport in S. pombe.
Kambe-Honjoh
H
H
Department of Agricultural Chemistry, University of Tokyo, Japan.
Yoda
K
K
Yamasaki
M
M
D001692:Biological Transport; D002463:Cell Membrane Permeability; D003600:Cytosol / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D008715:Methionine / Q000378:metabolism; D008854:Microscopy, Electron; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D012568:Schizosaccharomyces / Q000378:metabolism* / Q000648:ultrastructure; D013104:Spheroplasts / Q000378:metabolism / Q000648:ultrastructure; D013462:Sulfur Radioisotopes
D005656:Fungal Proteins; C048632:MF(ALPHA)1 protein, S cerevisiae; D011498:Protein Precursors; D029701:Saccharomyces cerevisiae Proteins; D013462:Sulfur Radioisotopes; D008715:Methionine