A novel hotspot of gelsolin instability triggers an alternative mechanism of amyloid aggregation

Bollati, Michela; Diomede, Luisa; Giorgino, Toni; Natale, Carmina; Fagnani, Elisa; Boniardi, Irene; Barbiroli, Alberto; Alemani, Rebecca; Beeg, Marten; Gobbi, Marco; Fakin, Ana; Mastrangelo, Eloise; Milani, Mario; Presciuttini, Gianluca; Gabellieri, Edi; Cioni, Patrizia; Rosa, Matteo de

doi:10.1016/j.csbj.2021.11.025

Cited by 3 publications

(5 citation statements)

References 65 publications

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“…Recombinant full-length GSN protein, carrying an N-terminal His 6 -tag, was produced as previously described. , Briefly, the human plasma isoform of GSN devoid of the signal peptide (mature form) was produced in Escherichia coli SHuffle cells (New England Biolabs) upon addition of 0.5 mM IPTG and incubation for 16 h at 18 °C. Cells were lysed in a Basic Z Bench top (Constant Systems Limited, U.K.) at 25 kPSI, and the clarified extract passed through a HisTrap HP column (all chromatographic media from GE-Healthcare).…”

Section: Theory and Methodsmentioning

confidence: 99%

“…In the absence of Ca 2+ , the actin binding sites are buried, limiting the ability of the GSN to interact with actin filaments. In this inactive state, GSN can be crystallized, but the resolution is relatively low and several stretches of the protein are too flexible to be modeled; such flexibility has been shown to be relevant for GSN physiopathology. − In this work, we have determined the ensemble of gelsolin structures in the closed and inactive state using SAXS data measured in the absence of calcium. Furthermore, as a second example of the applicability of hySAS, we refined a previously published protein–RNA complex.…”

Section: Introductionmentioning

confidence: 99%

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Accurate and Efficient SAXS/SANS Implementation Including Solvation Layer Effects Suitable for Molecular Simulations

Ballabio,

Paissoni,

Bollati

et al. 2023

J. Chem. Theory Comput.

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Small-angle X-ray and neutron scattering (SAXS/ SANS) provide valuable insights into the structure and dynamics of biomolecules in solution, complementing a wide range of structural techniques, including molecular dynamics simulations. As contrastbased methods, they are sensitive not only to structural properties but also to solvent−solute interactions. Their use in molecular dynamics simulations requires a forward model that should be as fast and accurate as possible. In this work, we demonstrate the feasibility of calculating SAXS and SANS intensities using a coarsegrained representation consisting of one bead per amino acid and three beads per nucleic acid, with form factors that can be corrected on the fly to account for solvation effects at no additional computational cost. By coupling this forward model with molecular dynamics simulations restrained with SAS data, it is possible to determine conformational ensembles or refine the structure and dynamics of proteins and nucleic acids in agreement with the experimental results. To assess the robustness of this approach, we applied it to gelsolin, for which we acquired SAXS data on its closed state, and to a UP1-microRNA complex, for which we used previously collected measurements. Our hybrid-resolution small-angle scattering (hySAS) implementation, being distributed in PLUMED, can be used with atomistic and coarse-grained simulations using diverse restraining strategies.

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Section: Theory and Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Accurate and Efficient SAXS/SANS Implementation Including Solvation Layer Effects Suitable for Molecular Simulations

Ballabio,

Paissoni,

Bollati

et al. 2023

J. Chem. Theory Comput.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Recombinant full-length GSN protein, carrying an N-terminal His 6 -tag, was produced as previously described. 50,71 Briefly, the human plasma isoform of GSN devoid of the signal peptide (mature form) was produced in Escherichia coli SHuffle cells (New England Biolabs) upon addition of 0.5 mM IPTG and incubation for 16 h at 18 °C. Cells were lysed in a Basic Z Bench top (Constant Systems Limited, U.K.) at 25 kPSI, and the clarified extract passed through a HisTrap HP column (all chromatographic media from GE-Healthcare).…”

Section: Bead Form Factor Parametrization and Validationmentioning

confidence: 99%

An Accurate and Efficient SAXS/SANS Implementation Including Solvation Layer Effects Suitable for Molecular Simulations.

Ballabio,

Paissoni,

Bollati

et al. 2023

Preprint

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Small-angle X-ray and neutron scattering (SAXS/SANS) provide valuable information on biomolecules in solution and are particularly well-suited to complement a wide range of structural techniques, including molecular dynamics simulations. As contrast-based techniques, they are sensitive not only to structural properties but also to the solvent-solute interactions. Their use in molecular dynamics simulations requires a forward model that should be as fast and accurate as possible. Here we show that it is possible to calculate SAXS and SANS using a coarse-grained representation of one bead per amino acid and three beads for nucleic acid with form factors that can be corrected on-the-fly to account for their interaction with the solvent at no additional computational cost. We use SAXS data measured for the closed state of Gelsolin and previous data for a UP1-microRNA complex to show that such approach allows to refine the structure and dynamics of proteins and nucleic acids. Our hybrid resolution small angle scattering (hySAS) implementation, being distributed in PLUMED, is ready to be coupled with atomistic and coarse-grained simulations using diverse restraining strategies.

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“…The ability of the LB peptidomimetics to reduce the toxicity of GAC 182-192 aggregates in vivo was investigated by employing the invertebrate C. elegans, a well-validated animal model able to recognize the toxic forms of amyloidogenic proteins, including different G2 and full-length GSN variants [22,23]. Thanks to their sensitivity to sublethal doses of chemical stressors, worms react to amyloid oligomers and/or aggregates by reducing the contraction and relaxation of the pharyngeal muscle, defined as "pumping rate", providing a direct relationship between the protein structure and its toxicity [37][38][39][40].…”

Section: Lb-5 and Lb-6 Counteract The Toxicity Of Aggregated Gac 182-...mentioning

confidence: 99%

“…We have recently demonstrated that the G4:G5-linked variants, namely A551P, E553K, and M517R, are not susceptible to furin proteolysis and are endowed with amyloid aggregation propensity in their full-length form [22]. Other yet-to-be-identified amyloidogenic stretches of the protein may be responsible for this proteolysis-independent aggregation pathway.…”

Section: Introductionmentioning

confidence: 99%

Rational Design of a Peptidomimetic Inhibitor of Gelsolin Amyloid Aggregation

Bollati

Peqini

Barone

et al. 2022

IJMS

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Gelsolin amyloidosis (AGel) is characterized by multiple systemic and ophthalmic features resulting from pathological tissue deposition of the gelsolin (GSN) protein. To date, no cure is available for the treatment of any form of AGel. More than ten single-point substitutions in the GSN gene are responsible for the occurrence of the disease and, among them, D187N/Y is the most widespread variant. These substitutions undergo an aberrant proteolytic cascade, producing aggregation-prone peptides of 5 and 8 kDa, containing the Gelsolin Amyloidogenic Core, spanning residues 182–192 (GAC182–192). Following a structure-based approach, we designed and synthesized three novel sequence-specific peptidomimetics (LB-5, LB-6, and LB-7) built on a piperidine-pyrrolidine unnatural amino acid. LB-5 and LB-6, but not LB-7, efficiently inhibit the aggregation of the GAC182–192 amyloidogenic peptides at sub-stoichiometric concentrations. These peptidomimetics resulted also effective in vivo, in a C. elegans-based assay, in counteracting the proteotoxicity of aggregated GAC182–192. These data pave the way to a novel pharmacological strategy against AGel and also validate a toolbox exploitable in other amyloidogenic diseases.

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A novel hotspot of gelsolin instability triggers an alternative mechanism of amyloid aggregation

Abstract: Graphical abstract

Cited by 3 publications

References 65 publications

Accurate and Efficient SAXS/SANS Implementation Including Solvation Layer Effects Suitable for Molecular Simulations

Accurate and Efficient SAXS/SANS Implementation Including Solvation Layer Effects Suitable for Molecular Simulations

An Accurate and Efficient SAXS/SANS Implementation Including Solvation Layer Effects Suitable for Molecular Simulations.

Rational Design of a Peptidomimetic Inhibitor of Gelsolin Amyloid Aggregation

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