The Bacillus licheniformis BlaP beta-lactamase as a model protein scaffold to study the insertion of protein fragments.
Using genetic engineering technologies, the chitin-binding domain (ChBD) of the human macrophage chitotriosidase has been inserted into the host protein BlaP, a class A beta-lactamase produced by Bacillus licheniformis. The product of this construction behaved as a soluble chimeric protein that conserves both the capacity to bind chitin and to hydrolyze beta-lactam moiety. Here we describe the biochemical and biophysical properties of this protein (BlaPChBD). This work contributes to a better understanding of the reciprocal structural and functional effects of the insertion on the host protein scaffold and the heterologous structured protein fragments. The use of BlaP as a protein carrier represents an efficient approach to the functional study of heterologous protein fragments.
Vandevenne
Marylène
M
Macromolécules Biologiques, Centre d'Ingénierie des Protéines, Université de Liège, Liège, Belgium.
Filee
Patrice
P
Scarafone
Natacha
N
Cloes
Benoît
B
Gaspard
Gilles
G
Yilmaz
Nursel
N
Dumoulin
Mireille
M
François
Jean-Marie
JM
Frère
Jean-Marie
JM
Galleni
Moreno
M
D001407:Bacillus / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002686:Chitin / Q000378:metabolism; D004795:Enzyme Stability; D006596:Hexosaminidases / Q000737:chemistry; D008958:Models, Molecular; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D001618:beta-Lactamases / Q000737:chemistry* / Q000235:genetics
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D002686:Chitin; D006596:Hexosaminidases; C086400:chitotriosidase; D001618:beta-Lactamases