Poster (Scientific congresses and symposiums)
Production of pure and functional Bone Morphognetic Protein-2
Robert, Charly; Matagne, André; Filée, Patrice et al.
201812th International BMP Conference
 

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Keywords :
BMP; Production; Bone morphogenetic protein
Abstract :
[en] Bone Morphognetic Protein-2 (BMP-2), which belongs to the transforming growth factor β (TGF-β) superfamily, plays an important role during bone regeneration and repair, and also during various stages of embryonic development. Noggin is a BMP-specific antagonist implicated in the regulation of BMP signaling pathways. Both BMP-2 and Noggin are dimeric molecules with each subunit containing several intramolecular disulfide bonds. This complex disulfide bonds network combined with the presence of large hydrophobic patches on the surface of these proteins cause their low solubility in aqueous solutions. As a result, BMP-2 and Noggin are commonly produced as inclusion bodies in Escherichia coli. The recovery of soluble and active molecules through in vitro refolding is difficult and the overall yield is low. Here we propose a new and simple method to produce biologically active BMP-2 and Noggin. Refolding of BMP-2 and Noggin was achieved in a one-step dilution and hydrophobic or ion exchange chromatography were used to purify the refolded proteins. The best conditions yielded significant quantities of dimeric BMP-2 and Noggin, with purity above 95%. Both BMP-2 and Noggin were characterized by optical spectroscopies and its biological activity was tested by measuring alkaline phosphatase activity in ATDC5 cells. These experiments indicated that our BMP-2 and Noggin display structural and functional properties similar to their commercial counterparts.
Research center :
CIP - Centre d'Ingénierie des Protéines - ULiège
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Robert, Charly ;  Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Matagne, André  ;  Université de Liège - ULiège > Département des sciences de la vie > Enzymologie et repliement des protéines
Filée, Patrice;  Centre d'Economie Rurale (Marloie) - CER
Bruck, France;  Centre d'Economie Rurale (Marloie) - CER
Alié, Christian;  Centre d'Economie Rurale (Marloie) - CER
Language :
English
Title :
Production of pure and functional Bone Morphognetic Protein-2
Alternative titles :
[fr] Production d'une Protéine de Morphogenèse Osseuse 2 pure et fonctionnelle
Publication date :
27 October 2018
Number of pages :
A0
Event name :
12th International BMP Conference
Event organizer :
Kohei Miyazono (The University of Tokyo)
Event place :
Tokyo, Japan
Event date :
du 24 octobre 2018 au 28 octobre 2018
By request :
Yes
Audience :
International
Name of the research project :
AUTOBMP2
Funders :
Service public de Wallonie : Direction générale opérationnelle de l'économie, de l'emploi et de la recherche - DG06
Available on ORBi :
since 05 November 2018

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