Vandevenne Marylène, Filee Patrice, Scarafone Natacha, Cloes Benoît, Gaspard Gilles, Yilmaz Nursel, Dumoulin Mireille, François Jean-Marie, Frère Jean-Marie, Galleni Moreno
Macromolécules Biologiques, Centre d'Ingénierie des Protéines, Université de Liège, Liège, Belgium.
Protein Sci. 2007 Oct;16(10):2260-71. doi: 10.1110/ps.072912407.
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.
利用基因工程技术，已将人类巨噬细胞几丁质酶的几丁质结合结构域（ChBD）插入宿主蛋白BlaP中，BlaP是地衣芽孢杆菌产生的A类β-内酰胺酶。这种构建产物表现为一种可溶性嵌合蛋白，既保留了结合几丁质的能力，又保留了水解β-内酰胺部分的能力。在此，我们描述了这种蛋白质（BlaPChBD）的生化和生物物理特性。这项工作有助于更好地理解插入对宿主蛋白支架和异源结构化蛋白片段的相互结构和功能影响。使用BlaP作为蛋白质载体代表了一种研究异源蛋白片段功能的有效方法。
