Some characteristics of hormone (pheromone) processing enzymes in yeast.
The KEX2 gene-encoded, membrane-bound Ca2+-dependent thiol endoproteinase, proteinase yscF, responsible for processing of the precursor protein of the sex pheromone alpha-factor of the yeast Saccharomyces cerevisiae was solubilized from the membraneous fraction and partially purified. Gel filtration revealed an apparent Mr of the native protein of around 150,000. Ca2+ concentration for half-maximal activity was in the micromolar range and concentration of the substrate Cbz-Tyr-Lys-Arg-4-nitroanilide for half-maximal velocity was 0.05 mM. The enzyme able to cleave basic amino acids from the carboxy-terminus of peptides and probably involved in final maturation of the alpha-factor peptides generated by proteinase yscF is membrane-associated, active at neutral pH and responds strongly to the serine proteinase inhibitor phenyl-methylsulfonyl fluoride as well as to -SH group blocking agents.
Wagner
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JC
Escher
C
C
Wolf
D H
DH
D002850:Chromatography, Gel; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005656:Fungal Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000072235:Mating Factor; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D043484:Proprotein Convertases; D011480:Protease Inhibitors; D011499:Protein Processing, Post-Translational; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D013381:Subtilisins
D005656:Fungal Proteins; D008565:Membrane Proteins; D010455:Peptides; D011480:Protease Inhibitors; D029701:Saccharomyces cerevisiae Proteins; D000072235:Mating Factor; D010450:Endopeptidases; C053108:carboxypeptidase ysc-alpha; D043484:Proprotein Convertases; D013381:Subtilisins; C058751:KEX2 protein, S cerevisiae