A similar protein portion for two exoglucanases secreted by Saccharomyces cerevisiae.
Exoglucanase (exo-1,3-beta-D-glucan glycohydrolase, EC 3.2.1.56) activity secreted by Saccharomyces cerevisiae into the culture medium was separated by ion exchange chromatography into two glycoprotein isoenzymes which contributed 10% (exoglucanase I) and 90% (exoglucanase II) towards the total activity. Analysis of the "in vitro" deglycosylated products by polyacrylamide gel electrophoresis under native or denaturing conditions indicated that the protein portions of both exoglucanases exhibited identical mobility, each one consisting of two polypeptides with Mr of 47,000 and 48,000. The same profile was shown by the exoglucanase secreted in the presence of tunicamycin. Antibodies raised against the protein portion of exoglucanase II did react with both native exoglucanases and their deglycosylated products with a pattern indicative of immunological identity. Digestion of the "in vitro" deglycosylated products of both exoglucanases with Staphylococcus aureus V-8 protease or trypsin generated the same proteolytic fragments in each case. Only exoglucanase II was secreted by protoplasts. These and previously reported results indicate that the protein portions of both isoenzymes may be the product of the same gene (or a family of related genes), and that exoglucanase I is a product of enzyme II, modified by a process occurring beyond the permeability barrier of the cell.
Ramírez
M
M
Departamento de Microbiología, Facultad de Ciencias, Universidad de Extremadura, Badajoz, Spain.
Hernández
L M
LM
Larriba
G
G
D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000032:analysis* / Q000302:isolation & purification; D043326:Glucan 1,3-beta-Glucosidase; D005959:Glucosidases / Q000032:analysis*; D005779:Immunodiffusion; D007527:Isoenzymes / Q000032:analysis* / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D001617:beta-Glucosidase / Q000032:analysis* / Q000302:isolation & purification
D005656:Fungal Proteins; D007527:Isoenzymes; D005959:Glucosidases; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase