title: Immobilized endoproteinase Glu-C to magnetic bead cellulose as a tool in proteomic analysis.
abstract: Magnetic bead cellulose activated with divinyl sulfone was used for the immobilization of Staphylococcus aureus endoproteinase Glu-C (EC 3.4.21.19). The immobilized proteinase was characterized by increased thermostability, by decreased self-cleavage activity, and a possibility of repeated use. The prepared immobilized enzyme was applied for the proteolytic cleavage of α-casein and BSA under different conditions (different composition of buffers, different pH, and different time of digestion). The possibilities of the direct use of enzyme reaction products for MALDI TOF MS analysis were shown.
authors.affiliation: Institute of Pathological Physiology, First Faculty of Medicine, Charles University, Prague, Czech Republic. petr.prikryl@lf1.cuni.cz
authors.forename: Petr
authors.initials: P
authors.lastname: Prikryl
authors.forename: Marie
authors.initials: M
authors.lastname: Ticha
authors.forename: Zdenka
authors.initials: Z
authors.lastname: Kucerova
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D002364:Caseins / Q000737:chemistry; D002482:Cellulose / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008280:Magnetics; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D012697:Serine Endopeptidases / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013211:Staphylococcus aureus / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D002364:Caseins; D004800:Enzymes, Immobilized; D012710:Serum Albumin, Bovine; D002482:Cellulose; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase