Insights on the emerging biotechnology of histidine-rich peptides

López‐Laguna, Hèctor; Voltà‐Durán, Eric; Parladé, Eloi; Villaverde, Antonio; Vázquez, Esther; Unzueta, Ugutz

doi:10.1016/j.biotechadv.2021.107817

Cited by 46 publications

(33 citation statements)

References 178 publications

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“…As expected for a H6-tagged protein [41,42], the addition of equimolar amounts of Zn 2+ to the monomeric protein versions promoted a shift in the DLS plots from 6.2-6.7 to 10.2-10.5 nm, indicative of protein assembling as NPs (Zn NPs, Fig. 2a).…”

Section: Resultssupporting

confidence: 64%

The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles

Voltà‐Durán

Sánchez²,

Hèctor-López-Laguna³

et al. 2022

Sci. China Mater.

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Histidine-rich peptides confer self-assembling properties to recombinant proteins through the supramolecular coordination with divalent cations. This fact allows the cost-effective, large-scale generation of microscopic and macroscopic protein materials with intriguing biomedical properties. Among such materials, resulting from the simple bioproduction of protein building blocks, homomeric nanoparticles are of special value as multivalent interactors and drug carriers. Interestingly, we have here identified that the assembly of a given His-tagged protein might render distinguishable categories of self-assembling protein nanoparticles. This fact has been scrutinized through the nanobody-containing fusion proteins EM1-GFP-H6 and A3C8-GFP-H6, whose biosynthesis results in two distinguishable populations of building blocks. In one of them, the assembling and disassembling is controllable by cations. However, a second population immediately self-assembles upon purification through a non-regulatable pathway, rendering larger nanoparticles with specific biological properties. The structural analyses of both model proteins and nanoparticles revealed important conformational variability in the building blocks. This fact renders different structural and functional categories of the final soft materials resulting from the participation of energetically unstable intermediates in the oligomerization process. These data illustrate the complexity of the Hismediated protein assembling in recombinant proteins but they also offer clues for a better design and refinement of protein-based nanomedicines, which, resulting from biological fabrication, show an architectonic flexibility unusual among biomaterials.

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Section: Resultssupporting

confidence: 64%

The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles

Voltà‐Durán

Sánchez²,

Hèctor-López-Laguna³

et al. 2022

Sci. China Mater.

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“…If His-rich peptides have a role in the physiological aggregation of recombinant proteins, IB formation and properties may be manipulated, a possibility that would expand the versatility of such natural materials. This is particularly important since His-rich peptides, such as the hexahistidine (H6), are very often used as purification tags of recombinant proteins and, therefore, present in most of the recombinant polypeptides produced in research and industry [ 43 ].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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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.

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“…The modular protein T22-GFP-H6 has been used in this study as a model. T22 (red box) is a cationic peptide targeting CXCR4 [53], L (brown box) is a peptidic linker (GGRSSRSS) that provides flexibility, GFP (green box) is an enhanced GFP and H6 is a hexahistidine tag (black box) [54,55]. H6 is useful for both chromatographic purification of the protein [54][55][56][57] and cation-mediated assembly [55,58].…”

Section: Resultsmentioning

confidence: 99%

Time-Prolonged Release of Tumor-Targeted Protein–MMAE Nanoconjugates from Implantable Hybrid Materials

et al. 2022

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The sustained release of small, tumor-targeted cytotoxic drugs is an unmet need in cancer therapies, which usually rely on punctual administration regimens of non-targeted drugs. Here, we have developed a novel concept of protein–drug nanoconjugates, which are packaged as slow-releasing chemically hybrid depots and sustain a prolonged secretion of the therapeutic agent. For this, we covalently attached hydrophobic molecules (including the antitumoral drug Monomethyl Auristatin E) to a protein targeting a tumoral cell surface marker abundant in several human neoplasias, namely the cytokine receptor CXCR4. By this, a controlled aggregation of the complex is achieved, resulting in mechanically stable protein–drug microparticles. These materials, which are mimetics of bacterial inclusion bodies and of mammalian secretory granules, allow the slow leakage of fully functional conjugates at the nanoscale, both in vitro and in vivo. Upon subcutaneous administration in a mouse model of human CXCR4+ lymphoma, the protein–drug depots release nanoconjugates for at least 10 days, which accumulate in the tumor with a potent antitumoral effect. The modification of scaffold cell-targeted proteins by hydrophobic drug conjugation is then shown as a novel transversal platform for the design of slow releasing protein–drug depots, with potential application in a broad spectrum of clinical settings.

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Insights on the emerging biotechnology of histidine-rich peptides

Cited by 46 publications

References 178 publications

The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles

The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles

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

Time-Prolonged Release of Tumor-Targeted Protein–MMAE Nanoconjugates from Implantable Hybrid Materials

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