Aggregation of gelsolin wild-type and G167K/R, N184K, and D187N/Y mutant peptides and inhibition

Ahmad, Mohanad; Esposto, Josephine; Golec, Camilla; Wu, Colin G.; Martic-Milne, Sanela

doi:10.1007/s11010-021-04085-6

Cited by 5 publications

(4 citation statements)

References 47 publications

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“…The relevance of these studies is questionable because the amyloidogenic fragment is still buried inside the folded domain. With regards to synthetic material, only short stretches of AGelD187N 173-242 have been used in aggregation studies [16,[18][19][20]. These short peptides are believed to represent the amyloidogenic region of the fragment.…”

Section: Introductionmentioning

confidence: 99%

Development of an in vitro aggregation assay for long synthetic polypeptide, amyloidogenic gelsolin fragment AGelD187N 173–242

Leimu,

Haavisto,

Nesati

et al. 2023

PLoS ONE

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Aggregation of the gelsolin protein fragment is the hallmark of the hereditary systemic disease gelsolin amyloidosis. As with other protein misfolding diseases, there is an urgent need for efficient disease-modifying treatment for gelsolin amyloidosis. The formation of amyloids can be reproduced by incubating the disease-causing amyloidogenic 8 kDa polypeptide, 70-residue gelsolin protein fragment, AGelD187N 173–242, in vitro and monitoring the process by thioflavin T dye. However, for screening of potential aggregation inhibitors, the required protein amounts are large and the biotechnological production of amyloidogenic proteins has many challenges. Conversely, use of shorter synthetic regions of AGelD187N 173–242 does not mimic the in vivo aggregation kinetics of full-length fragment as they have different aggregation propensity. In this study, we present an in vitro aggregation assay for full-length AGelD187N 173–242 that has been produced by solid-phase chemical synthesis and after that monomerized carefully. Chemical synthesis allows us to produce high quantities of full-length fragment efficiently and at low cost. We demonstrate that the generated aggregates are fibrillar in nature and how the purity, terminal modification, initial aggregates and seeding affect the aggregation kinetics of a synthetic gelsolin fragment. We also present sufficient quality criteria for the initial monomerized synthetic polypeptide.

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Section: Introductionmentioning

confidence: 99%

Development of an in vitro aggregation assay for long synthetic polypeptide, amyloidogenic gelsolin fragment AGelD187N 173–242

Leimu,

Haavisto,

Nesati

et al. 2023

PLoS ONE

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“…The larger product of furin activity, the C68 fragment, undergoes another proteolytic event that eventually leads to the production of two aggregation-prone peptides of 5 and 8 kDa [17]. These peptides readily aggregate because they contain an amyloidogenic core sequence, called Gelsolin Amyloidogenic Core (GAC), identified in systematic studies as spanning residues 182 to 192 [18] or 187 to 193 [19] of GSN. Other larger analogs harbouring GAC, such as a 15-mer [20] and the isolated G2 domain [21], were shown to recapitulate pathological GSN aggregation in vitro.…”

Section: Introductionmentioning

confidence: 99%

“…The inhibition of the aggregation of GSN in the form of aggregation-prone fragments or the full length, to prevent the deposition of large aggregates and the circulation of highly toxic soluble oligomers can thus be targeted to slow down or even block pathological aggregation of GSN. A few small molecules such as non-selective polyphenols, phospholipids, and other repurposed drugs [19,20,25] were shown to modulate GSN aggregation in vitro and counter the associated toxicity.…”

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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“…Amyloid fibrils can deposit as amyloid plaques in various tissues throughout the body, resulting in a wide range of pathological conditions [ 3 ]. Point mutations of amyloidogenic proteins (Aβ [ 4 ], transthyretin [ 5 ], apolipoprotein [ 6 ], gelsolin [ 7 ], and synuclein [ 8 ]) are known to be responsible for several familial amyloid disorders. These mutations can provoke protein aggregation by changing the protein’s physico-chemical properties or the stability of the native structure via changes in charge network, hydrophobicity, and propensity to form an intermolecular β-sheet structure [ 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

Pathogenic D76N Variant of β2-Microglobulin: Synergy of Diverse Effects in Both the Native and Amyloid States

Bulyáki

Kun

Molnár

et al. 2021

Biology

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β2-microglobulin (β2m), the light chain of the MHC-I complex, is associated with dialysis-related amyloidosis (DRA). Recently, a hereditary systemic amyloidosis was discovered, caused by a naturally occurring D76N β2m variant, which showed a structure remarkably similar to the wild-type (WT) protein, albeit with decreased thermodynamic stability and increased amyloidogenicity. Here, we investigated the role of the D76N mutation in the amyloid formation of β2m by point mutations affecting the Asp76-Lys41 ion-pair of WT β2m and the charge cluster on Asp38. Using a variety of biophysical techniques, we investigated the conformational stability and partial unfolding of the native state of the variants, as well as their amyloidogenic propensity and the stability of amyloid fibrils under various conditions. Furthermore, we studied the intermolecular interactions of WT and mutant proteins with various binding partners that might have in vivo relevance. We found that, relative to WT β2m, the exceptional amyloidogenicity of the pathogenic D76N β2m variant is realized by the deleterious synergy of diverse effects of destabilized native structure, higher sensitivity to negatively charged amphiphilic molecules (e.g., lipids) and polyphosphate, more effective fibril nucleation, higher conformational stability of fibrils, and elevated affinity for extracellular components, including extracellular matrix proteins.

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Aggregation of gelsolin wild-type and G167K/R, N184K, and D187N/Y mutant peptides and inhibition

Cited by 5 publications

References 47 publications

Development of an in vitro aggregation assay for long synthetic polypeptide, amyloidogenic gelsolin fragment AGelD187N 173–242

Development of an in vitro aggregation assay for long synthetic polypeptide, amyloidogenic gelsolin fragment AGelD187N 173–242

Rational Design of a Peptidomimetic Inhibitor of Gelsolin Amyloid Aggregation

Pathogenic D76N Variant of β2-Microglobulin: Synergy of Diverse Effects in Both the Native and Amyloid States

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