The Poly-Histidine Tag H6 Mediates Structural and Functional Properties of Disintegrating, Protein-Releasing Inclusion Bodies

• Main Authors: Julieta María Sánchez (Author), José Vicente Carratalá (Author), Naroa Serna (Author), Ugutz Unzueta (Author), Verónica Nolan (Author), Alejandro Sánchez-Chardi (Author), Eric Voltà-Durán (Author), Hèctor López-Laguna (Author), Neus Ferrer-Miralles (Author), Antonio Villaverde (Author), Esther Vazquez (Author)
• Format: Book
• Published: MDPI AG, 2022-03-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.
• Notes: 10.3390/pharmaceutics14030602; 1999-4923
• Keywords: inclusion bodies; functional amyloids; protein secretion; His-cation coordination; biomaterials; protein materials; Pharmacy and materia medica; RS1-441
• Source: Pharmaceutics, Vol 14, Iss 3, p 602 (2022)

The coordination between histidine-rich peptides and divalent cations supports the formation of nano- and micro-scale protein biomaterials, including toxic and non-toxic functional amyloids, which can be adapted as drug delivery systems. Among them, inclusion bodies (IBs) formed in recombinant bacteria have shown promise as protein depots for time-sustained protein release. We have demonstrated here that the hexahistidine (H6) tag, fused to recombinant proteins, impacts both on the formation of bacterial IBs and on the conformation of the IB-forming protein, which shows a higher content of cross-beta intermolecular interactions in H6-tagged versions. Additionally, the addition of EDTA during the spontaneous disintegration of isolated IBs largely affects the protein leakage rate, again protein release being stimulated in His-tagged materials. This event depends on the number of His residues but irrespective of the location of the tag in the protein, as it occurs in either C-tagged or N-tagged proteins. The architectonic role of H6 in the formation of bacterial IBs, probably through coordination with divalent cations, offers an easy approach to manipulate protein leakage and to tailor the applicability of this material as a secretory amyloidal depot in different biomedical interfaces. In addition, the findings also offer a model to finely investigate, in a simple set-up, the mechanics of protein release from functional secretory amyloids.