The Bacillus licheniformis BlaP beta-lactamase as a model protein scaffold to study the insertion of protein fragments.

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

Volume :

Issue :

Pages :

2260-71

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 19 November 2010

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