Unexpected influence of a C-terminal-fused His-tag on the processing of an enzyme and on the kinetic and folding parameters

• Published in: FEBS letters Vol. 413; no. 2; pp. 194 - 196
• Main Authors: Ledent, Philippe, Duez, Colette, Vanhove, Marc, Lejeune, Annabelle, Fonzé, Eveline, Charlier, Paulette, Rhazi-Filali, Fouzia, Thamm, Iris, Guillaume, Gilliane, Samyn, Bart, Devreese, Bart, Van Beeumen, Jozef, Lamotte-Brasseur, Josette, Frère, Jean-Marie
• Format: Journal Article Web Resource
• Language: English
• Published: England Elsevier B.V 18.08.1997; Elsevier Science
• Subjects: Bacillus; Bacillus - enzymology; beta-Lactamases - chemistry; beta-Lactamases - genetics; beta-Lactamases - isolation & purification; beta-Lactamases - metabolism; Biochemistry; Biochemistry, biophysics & molecular biology; Biochimie, biophysique & biologie moléculaire; biophysics; Crystallography, X-Ray; His-tag; Histidine - metabolism; Kinetics; Life sciences; Membrane Proteins; molecular biology; Peptides - metabolism; Protein Folding; Protein Processing, Post-Translational; Protein Sorting Signals - metabolism; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; Sciences du vivant; Serine Endopeptidases - metabolism; Thermophile; β-Lactamase; β-Lactamase; Thermophile; His-tag; Bacillus
• ISSN: 0014-5793; 1873-3468; 1873-3468
• DOI: 10.1016/S0014-5793(97)00908-3

The addition of a poly-His C-terminal extension, designed to facilitate the purification of the protein, to the β-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.