Expression, purification and functional characterization of Wnt signaling co-receptors LRP5 and LRP6

被引：5

作者：

Fleury, Damien ^{[1
]}

Vayssiere, Beatrice ^{[2
]}

Touitou, Robert ^{[1
]}

Gillard, Corinne ^{[1
]}

Lebhar, Helene ^{[1
]}

Rawadi, Georges ^{[2
]}

Mollat, Patrick ^{[1
]}

机构：

[1] Galapagos, Prot Sci, F-93230 Romainville, France

[2] Galapagos, Translat Sci Labs, F-93230 Romainville, France

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2010年 / 70卷 / 01期

关键词：

LRP5/6; Wnt pathway; Dkk proteins; Purification; Characterization; Binding;

D O I：

10.1016/j.pep.2009.08.012

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Activation of the Writ signaling cascade plays a pivotal role during development and in various disease states. Wnt signals are transduced by seven-transmembrane Frizzled (Fz) proteins and the single-transmembrane LDL-receptor-related proteins 5 or 6 (LRP5/6). Genetic mutations resulting in a loss or gain of function of LRP5 in humans lead to osteopenia and bone formation, respectively. These findings demonstrate the genetic link between LRP5 signaling and the regeneration of bone mass. Herein we describe for the first time the production and characterization of soluble ectodomains of LRP5 and LRP6, (EC-LRP5, EC-LRP6). We have produced these proteins in amounts that are compatible with both in vitro and cell-based assays to study their binding properties. Purified EC-LRP5 and EC-LRP6 were able to interact with Wnt signaling components Dkk1 and Dkk2 and their functionality was confirmed in cell-based Wnt signaling assays. Hence, tools are now available to explore LRP5/6 interaction with other proteins and to screen for synthetic or natural compounds and biologics that might be novel therapeutics targeting the Wnt pathway. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：39 / 47

页数：9

共 35 条

[1] Correspondence - A colipase fold in the carboxy-terminal domain of the Wnt antagonists - the Dickkopfs [J].