Bacillus/enzymology; Bacterial Proteins/chemistry/genetics; Chitin/metabolism; Enzyme Stability; Hexosaminidases/chemistry; Models, Molecular; Protein Denaturation; Protein Structure, Tertiary; Recombinant Fusion Proteins/chemistry; beta-Lactamases/chemistry/genetics
Abstract :
[en] 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.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Vandevenne, Marylène ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Filée, Patrice ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Scarafone, Natacha ; Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Cloes, Benoit
Gaspard, Gilles
Yilmaz, Nursel
Dumoulin, Mireille ; Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
François, Jean-Marie ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Frère, Jean-Marie ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Galleni, Moreno ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques
Language :
English
Title :
The Bacillus licheniformis BlaP beta-lactamase as a model protein scaffold to study the insertion of protein fragments.
Publication date :
2007
Journal title :
Protein Science: A Publication of the Protein Society
ISSN :
0961-8368
eISSN :
1469-896X
Publisher :
Cold Spring Harbor Laboratory Press, Woodbury, United States - New York
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