title: Rapid analysis of enzymatic digests of a bacterial protease of the subtilisin type and a "bio-engineered" variant by high-performance liquid chromatography-frit fast atom bombardment mass spectrometry.
abstract: Amino acid sequencing of a subtilisin-type bacterial protease and a bio-engineered variant was carried out by investigating various enzymatic digests using HPLC-frit fast atom bombardment MS methods. The fast atom bombardment mass spectral data allowed rapid identification of the enzymatically generated peptides and differentiation between both proteins. The feasibility of determining the positions and nature of mutations in the amino acid sequence depends mainly on the size of the peptides containing the modifications.
authors.affiliation: Bijvoet Centre for Biomolecular Research, Department of Mass Spectrometry, Utrecht University, Netherlands.
authors.forename: W D
authors.initials: WD
authors.lastname: van Dongen
authors.forename: C
authors.initials: C
authors.lastname: Versluis
authors.forename: P D
authors.initials: PD
authors.lastname: van Wassenaar
authors.forename: C G
authors.initials: CG
authors.lastname: de Koster
authors.forename: W
authors.initials: W
authors.lastname: Heerma
authors.forename: J
authors.initials: J
authors.lastname: Haverkamp
mesh_terms: D000595:Amino Acid Sequence; D001329:Autolysis; D001419:Bacteria / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D002918:Chymotrypsin; D003488:Cyanogen Bromide; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010434:Pepsin A; D010446:Peptide Fragments / Q000032:analysis; D011994:Recombinant Proteins / Q000032:analysis; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013381:Subtilisins / Q000032:analysis*; D014357:Trypsin
chemical_list: D010446:Peptide Fragments; D011994:Recombinant Proteins; D013381:Subtilisins; D002918:Chymotrypsin; D014357:Trypsin; D010434:Pepsin A; D003488:Cyanogen Bromide