Chaperone-mediated production of active homodimer human bone morphogenetic protein – 2 in E. coli

• Published in: Protein expression and purification 2023-06, Vol.206, p.106245-106245, Article 106245
• Main Authors: Lykoshin, Dmitry D., Kostromina, Maria A., Azmukova, Veronika R., Esipov, Roman S.
• Format: Article
• Language: English
• Subjects: Bone Morphogenetic Protein 2 - chemistry; Bone Morphogenetic Protein 7 - metabolism; Escherichia coli - genetics; Escherichia coli - metabolism; Homodimeric protein; Humans; Molecular Chaperones - genetics; Molecular Chaperones - metabolism; Osteogenesis; Osteogenic activity; Protein purification; Recombinant human bone morphogenetic protein (rhBMP-2); Recombinant Proteins - chemistry; Soluble expression
• ISSN: 1046-5928; 1096-0279
• DOI: 10.1016/j.pep.2023.106245

Human bone morphogenetic protein 2 (hBMP-2) plays a leading role in the process of osteogenesis and is one of the key components of osteoplastic materials, ensuring their high osteoinduction. In order to obtain a homodimeric form hBMP-2 using the E. coli expression system, a number of problems associated with refolding in vitro and purification from monomer and oligomeric forms must be solved. The developed method for co-expression of the target protein with chaperone proteins makes it possible to obtain the biologically active homodimeric form of hBMP-2 in vivo. Purification with simple ion-exchange sorbents without the use of denaturing reagents affecting the structure of the protein molecule provides a chromatographic purity of the product of at least 97%. The expressed hBMP-2 was identified by Western blotting and the LC-ESI-TOF mass spectrometry confirmed its molecular weight of 26052.72 Da. Circular dichroism spectroscopy showed that recombinant hBMP-2 has a native secondary structure. •Co-expression of the BMP-2 with GroEL-GroES-TF chaperone protein complex.•Soluble homodimeric form of tag-free BMP-2 used E.coli expression system.•High yield and native structure of BMP-2 were obtained in E. coli.