title: Activity-based identification of secreted serine proteases of the filamentous fungus, Ophiostoma.
abstract: A general activity probe was synthesized and applied to the supernatant of a filamentous fungus, Ophiostoma, culture to identify directly the secreted serine proteases by covalent enzyme labeling. The activity probe contained a chemically reactive group that reacted with, and thus covalently labeled, the serine residues of only active proteases and not heat-inactivated proteases. The activity probe also contained a fluorescent group that allowed for the subsequent visualization of the captured proteases in 1-D gels and their identification by N-terminal sequencing. This use of a chemical probe led to the rapid discovery of subtilisin-like serine protease of Ophiostoma. Two hypothetical proteins were also captured, with one being a probable endopeptidase K type of protease.
authors: Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, NSW, Australia.
authors: Caiyan
authors: C
authors: Wu
authors: Qiang
authors: Q
authors: Xu
authors: Fei
authors: F
authors: Liu
authors: K M Helena
authors: KM
authors: Nevalainen
mesh_terms: D001203:Ascomycota / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015394:Molecular Structure; D020539:Sequence Analysis, Protein; D012694:Serine / Q000737:chemistry / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D020860:Subtilisin / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D005656:Fungal Proteins; D012694:Serine; D012697:Serine Endopeptidases; D020860:Subtilisin