title: Unexpected influence of a C-terminal-fused His-tag on the processing of an enzyme and on the kinetic and folding parameters.
abstract: The addition of a poly-His C-terminal extension, designed to facilitate the purification of the protein, to the beta-lactamase of a thermophilic Bacillus licheniformis strain modified the site of action of the signal peptidase. This resulted in the secretion of a protein with a different N-terminus, showing that this type of protein engineering might not always be as 'neutral' as generally assumed.
authors.affiliation: Centre for Protein Engineering, Université de Liège, Institut de Chimie, Belgium.
authors.forename: P
authors.initials: P
authors.lastname: Ledent
authors.forename: C
authors.initials: C
authors.lastname: Duez
authors.forename: M
authors.initials: M
authors.lastname: Vanhove
authors.forename: A
authors.initials: A
authors.lastname: Lejeune
authors.forename: E
authors.initials: E
authors.lastname: Fonzé
authors.forename: P
authors.initials: P
authors.lastname: Charlier
authors.forename: F
authors.initials: F
authors.lastname: Rhazi-Filali
authors.forename: I
authors.initials: I
authors.lastname: Thamm
authors.forename: G
authors.initials: G
authors.lastname: Guillaume
authors.forename: B
authors.initials: B
authors.lastname: Samyn
authors.forename: B
authors.initials: B
authors.lastname: Devreese
authors.forename: J
authors.initials: J
authors.lastname: Van Beeumen
authors.forename: J
authors.initials: J
authors.lastname: Lamotte-Brasseur
authors.forename: J M
authors.initials: JM
authors.lastname: Frère
mesh_terms: D001407:Bacillus / Q000201:enzymology; D018360:Crystallography, X-Ray; D006639:Histidine / Q000378:metabolism; D007700:Kinetics; D008565:Membrane Proteins; D010455:Peptides / Q000378:metabolism*; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D001618:beta-Lactamases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C033223:polyhistidine; D006639:Histidine; D012697:Serine Endopeptidases; C026515:type I signal peptidase; D001618:beta-Lactamases