Role of the C-terminal cysteines in virus-like particle formation and oligomerization of the hepatitis E virus ORF2 truncated proteins

Liu, Zhenzhen; Behloul, Nouredine; Baha, Sarra; Wei, Wenjuan; Tao, Wanru; Zhang, Tingying; Li, Wei; Shi, Ruihua; Meng, Jingjing

doi:10.1016/j.virol.2020.01.011

Cited by 7 publications

(1 citation statement)

References 33 publications

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“…On the other hand, three cysteine residues are mechanistically identified as the minimal number envisaging the generation of oligomers beyond mere dimers and, also, as an adequate value to avoid complex multivalent network linking. Such a "three" value was supported by previous studies on hepatitis E virus-like particles (VLPs) 48 and modified green fluorescent proteins (GFPs), 49,50 in which their self-assembling properties were ablated when the number of Cys residues in the cross-interactive stretches was reduced below three. Therefore, by combining these concepts, a His−Cys hybrid tag, which contains such a minimum number of cysteines, was designed as HHHCHCHCH (H3C), by alternating those three cysteine residues starting from the C-terminal of the H6 tag sequence.…”

Section: ■ Results and Discussionmentioning

confidence: 87%

Biofabrication of Self-Assembling Covalent Protein Nanoparticles through Histidine-Templated Cysteine Coupling

López‐Laguna

Rueda

Martínez-Torró

et al. 2023

ACS Sustainable Chem. Eng.

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Nanoscale protein materials show increasing applications in biotechnology and biomedicine, addressing catalysis, drug delivery, or tissue engineering. Although protein oligomerization is reachable through several engineering approaches, including the use of divalent cations for histidine-rich stretches, the effectiveness of cation-His binding is influenced by protein conformation, media composition, and chelating agents. Thus, looking for powerful, green, cross-linker-free, and transversal oligomerization platforms, we have built a histidine-templated cysteine-coupling concept. On this basis, we have engineered a Cys-containing, H6-derived His−Cys hybrid tag that enables the spontaneous and efficient self-assembling of tagged proteins into monodisperse nanoparticles through a highly ordered covalent binding process. Although the generated nanostructures are supported by disulfide bridge formation and exclusively reversed by reducing agents but not by chelating agents, the presence of cysteine residues does not disrupt the metal-binding abilities of histidine residues within the tag. This fact allows one to combine the one-step IMAC-based protein purification and, also, the Zn 2+ -induced formation of higher-order microparticulate materials as nanoparticle-releasing protein-only depots. The dual mode of cross-molecular interactivity shown by the hybrid tag and the structural robustness and stability of the resulting nanoparticles offer wide applicability of the green biofabrication concept proposed here for the further development of clinically usable protein materials.

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Section: ■ Results and Discussionmentioning

confidence: 87%