Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, 117997, Russia.
Protein Expr Purif. 2023 Jun;206:106245. doi: 10.1016/j.pep.2023.106245. Epub 2023 Feb 16.
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.
人骨形态发生蛋白 2（hBMP-2）在成骨过程中起主导作用，是成骨材料的关键成分之一，可确保其具有较高的成骨诱导活性。为了使用大肠杆菌表达系统获得同源二聚体形式的 hBMP-2，必须解决体外重折叠以及从单体和低聚体形式中进行纯化等相关问题。通过与伴侣蛋白共表达的方法，可以在体内获得具有生物活性的 hBMP-2 同源二聚体形式。使用简单的离子交换吸附剂进行纯化，而不使用影响蛋白质分子结构的变性试剂，可使产物的色谱纯度至少达到 97%。通过 Western blot 鉴定表达的 hBMP-2，并通过 LC-ESI-TOF 质谱确认其分子量为 26052.72 Da。圆二色光谱表明重组 hBMP-2 具有天然的二级结构。
