Production of recombinant human ameloblastin by a fully native purification pathway

0558955 - ÚOCHB 2023 RIV US eng J - Journal Article
Vetýšková, Veronika - Zouharová, Monika - Boušová, Kristýna
Production of recombinant human ameloblastin by a fully native purification pathway.
Protein Expression and Purification. Roč. 198, October (2022), č. článku 106133. ISSN 1046-5928. E-ISSN 1096-0279
R&D Projects: GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : ameloblastin * oligomerization * protein native conditions * purification * twin strep-tag
OECD category: Biochemistry and molecular biology
Impact factor: 1.6, year: 2022
Method of publishing: Limited access
https://doi.org/10.1016/j.pep.2022.106133

Ameloblastin (Ambn) is an intrinsically disordered protein (IDP) with a specific function of forming heterogenous homooligomers. The oligomeric function is led through a specific sequence encoded by exon 5 of Ambn. Due to the IDP character of Ambn to form oligomers, protein purification is subject to many challenges. Human ameloblastin (AMBN) and its two isoforms, I and II have already been purified as a recombinant protein in a bacterial expression system and functionally characterized in vitro. However, here we present a new purification protocol for the production of native AMBN in its original formation as a homooligomeric heterogeneous IDP. The purification process consists of three chromatographic steps utilizing His-tag and Twin Strep-tag affinity chromatography, along with size exclusion and reverse affinity chromatography. The presented workflow offers the production of AMBN in sufficient yield for in vitro protein characterizations and can be used to produce both AMBN isoforms I and II.
Permanent Link: https://hdl.handle.net/11104/0332436