The Poly-Histidine Tag H6 Mediates Structural and Functional Properties of Disintegrating, Protein-Releasing Inclusion Bodies
Por:
Sanchez, JM, Carratala, JV, Serna, N, Unzueta, U, Nolan, V, Sanchez-Chardi, A, Volta-Duran, E, Lopez-Laguna, H, Ferrer-Miralles, N, Villaverde, A, Vazquez, E
Publicada:
1 mar 2022
Resumen:
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.
Filiaciones:
Sanchez, JM:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Univ Nacl Cordoba, Inst Invest Biol & Tecnol IIBYT, ICTA, CONICET, Av Velez Sarsfield 1611,X 5016GCA, Cordoba, Argentina
UNC, Catedra Quim Biol, Dept Quim, FCEFyN, Av Velez Sarsfield 1611,X 5016GCA, Cordoba, Argentina
Carratala, JV:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Serna, N:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Univ Autonoma Barcelona, Nanoligent SL, Edifici Eureka, Barcelona 08193, Spain
Unzueta, U:
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Biomed Res Inst St Pau IIB St Pau, St Antoni Maria Claret 167, Barcelona 08025, Spain
Josep Carreras Leukaemia Res Inst, Barcelona 08025, Spain
Nolan, V:
Univ Nacl Cordoba, Inst Invest Biol & Tecnol IIBYT, ICTA, CONICET, Av Velez Sarsfield 1611,X 5016GCA, Cordoba, Argentina
UNC, Catedra Quim Biol, Dept Quim, FCEFyN, Av Velez Sarsfield 1611,X 5016GCA, Cordoba, Argentina
Sanchez-Chardi, A:
Univ Autonoma Barcelona, Serv Microscopia, Placa Civica S-N, Barcelona 08193, Spain
Univ Barcelona, Fac Biol, Dept Biol Evolut Ecol & Ciencies Ambientals, Av Diagonal 643, Barcelona 08028, Spain
Volta-Duran, E:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Lopez-Laguna, H:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Ferrer-Miralles, N:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Villaverde, A:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Vazquez, E:
Univ Autonoma Barcelona, Inst Biotecnol & Biomed, Placa Civica S-N, Barcelona 08193, Spain
Univ Autonoma Barcelona, Dept Genet & Microbiol, Placa Civica S-N, Barcelona 08193, Spain
CIBER Bioingn Biomat & Nanomed CIBER BBN, C Monforte Lemos 3-5, Madrid 28029, Spain
Green Published, gold, All Open Access, Gold, Green
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