Unexpected influence of a C-terminal-fused His-tag on the processing of an enzyme and on the kinetic and folding parameters.
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.
Ledent
P
P
Centre for Protein Engineering, Université de Liège, Institut de Chimie, Belgium.
Duez
C
C
Vanhove
M
M
Lejeune
A
A
Fonzé
E
E
Charlier
P
P
Rhazi-Filali
F
F
Thamm
I
I
Guillaume
G
G
Samyn
B
B
Devreese
B
B
Van Beeumen
J
J
Lamotte-Brasseur
J
J
Frère
J M
JM
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*
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