TY - JOUR
T1 - The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles.
AU - López Laguna, Héctor
AU - Villaverde Corrales, Antonio Pedro
AU - Voltà Durán, Eric
AU - Vazquez Gomez, Esther
AU - Sanchez ., Julieta Maria
AU - Sánchez Garcia, Laura
AU - de Marco, Ario
AU - Alejandro, Sánchez-Chardi
AU - Parladé, Eloi
AU - Unzueta Elorza, Ugutz
N1 - Funding Information:
We are indebted to the Agencia Estatal de Investigación (AEI) and to the Fondo Europeo de Desarrollo Regional (FED-ER) (PID2020-116174RB-I00, AEI/FEDER, UE). The AGAUR (2017SGR-229) and the CIBER-BBN (project NANOPROTHER) are granted to Villaverde A. Vázquez E received support from the AEI (PID2019-105416RB-I00/AEI/10.13039/501100011033 and CIBER-BBN (project NANOREMOTE). We also appreciate the support from the Javna Agencija za Raziskovalno dejavnost Republike Slovenije (ARRS/N4-0046 and ARRS/J4-9322) to de Marco A. Sánchez-García L was supported by a predoctoral fellowship from the AGAUR (2018FI_B2_00051), Voltà-Durán E by a predoctoral fellowship from Ministerio de Ciencia, Innovacion y Universidades (FPU18/04615), Unzueta U was supported by Miguel Servet contract (CP19/00028) from ISCIII co-funded by European Social Fund (ESF investing in your future) and by an ISCIII project (PI20/00400) co-funding FEDER (A way to make Europe). López-Laguna H was supported by a predoctoral fellowship from the AGAUR (2019FI_B00352). Villaverde A received an ICREA ACADEMIA award. Protein production was partially performed by the ICTS “NAN-BIOSIS”, more specifically by the Protein Production Platform of CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN)/IBB, at the UAB ( http://www.nanbiosis.es/portfolio/u1-protein-production-platform-ppp/ ). Cell culture experiments were performed at SCAC facilities in the UAB. Molecular graphics and analyses were performed with UCSF Chimera, developed by the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco, with support from NIH P41-GM103311. Electron microscopy studies were performed by the Servei de Microscòpia in the UAB.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/1/17
Y1 - 2022/1/17
N2 - 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.
AB - 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.
KW - CONFORMATIONAL QUALITY
KW - DIVALENT-CATIONS
KW - EXPRESSION
KW - PURIFICATION
KW - SPECTROSCOPY
KW - STABILITY
KW - biomaterials
KW - nanoparticles
KW - protein materials
KW - recombinant proteins
KW - self-assembling
UR - http://www.scopus.com/inward/record.url?scp=85124765987&partnerID=8YFLogxK
UR - https://www.mendeley.com/catalogue/c2c49fd6-97ad-3edf-91e6-5cb693dde134/
U2 - 10.1007/s40843-021-1914-0
DO - 10.1007/s40843-021-1914-0
M3 - Article
SN - 2095-8226
VL - 65
SP - 1662
EP - 1670
JO - Science China Materials
JF - Science China Materials
IS - 6
ER -