The Tempered Polymerization of Human Neuroserpin

Noto, Rosina; Santangelo, M.; Ricagno, Stéfano; Mangione, Maria Rosalia; Levantino, Matteo; Pezzullo, Margherita; Martorana, Vincenzo; Cupane, Antonio; Bolognesi, Martino; Manno, Mauro

doi:10.1371/journal.pone.0032444

Cited by 28 publications

(54 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The use of a compressed exponential law is not based on any particular model for supra-molecular assembly. However, it may indicate that the observed aggregation process involves different phases whose reaction rates change as a function of time [32], [33]. Possible scenarios could be determined by changes in the reactivity of the growing species in solution.…”

Section: Resultsmentioning

confidence: 99%

Unlocked Concanavalin A Forms Amyloid-like Fibrils from Coagulation of Long-lived “Crinkled” Intermediates

et al. 2013

View full text Add to dashboard Cite

Understanding the early events during amyloid aggregation processes is crucial to single out the involved molecular mechanisms and for designing ad hoc strategies to prevent and reverse amyloidogenic disorders. Here, we show that, in conditions in which the protein is positively charged and its conformational flexibility is enhanced, Concanavalin A leads to fibril formation via a non-conventional aggregation pathway. Using a combination of light scattering, circular dichroism, small angle X-ray scattering, intrinsic (Tryptophan) and extrinsic (ANS) fluorescence and confocal and 2-photon fluorescence microscopy we characterize the aggregation process as a function of the temperature. We highlight a multi-step pathway with the formation of an on-pathway long-lived intermediate and a subsequent coagulation of such “crinkled” precursors into amyloid-like fibrils. The process results in a temperature-dependent aggregation-coagulation pathway, with the late phase of coagulation determined by the interplay between hydrophobic and electrostatic forces. Our data provide evidence for the complex aggregation pathway for a protein with a highly flexible native conformation. We demonstrate the possibility to generate a long-lived intermediate whose proportion and occurrence are easily tunable by experimental parameters (i.e. temperature). As a consequence, in the case of aggregation processes developing through well-defined energy barriers, our results can open the way to new strategies to induce more stable in vitro on-pathway intermediate species through a minute change in the initial conformational flexibility of the protein. This will allow isolating and experimentally studying such transient species, often indicated as relevant in neurodegenerative diseases, both in terms of structural and cytotoxic properties.

show abstract

Section: Resultsmentioning

confidence: 99%

Unlocked Concanavalin A Forms Amyloid-like Fibrils from Coagulation of Long-lived “Crinkled” Intermediates

et al. 2013

View full text Add to dashboard Cite

show abstract

“…In recent studies, we showed how the structural details of dysfunctional NS conformers, polymer and latent NS, depend upon thermodynamic and environmental conditions [22,23] , in keeping with other studies [24] . Further, we proposed that the mechanism of NS polymerization is rate limited by the formation of an intermediate conformation prone to dimerization [25,26] , as in other serpins [27] , and is controlled by a peculiar link between depolymerization and concomitant latentization [25] . Further, Onda and coworkers have recently identified a refolding intermediate leading to the formation of polymers alike those formed by native NS [28] .…”

Section: Introductionmentioning

confidence: 99%

“…Beyond its relevance in human pathology, NS is an excellent model for biophysical studies of serpin polymerization due to its close structural homology with the archetypal serpin α1-antitrypsin (AAT) [15] , but also in view of its capability of forming polymers and latent conformers under thermal stress [8–10,22–26] or under acidic conditions [24] , even in the wild-type form. Indeed, NS is relatively less stable than AAT, as marked by the achievement of fragile NS oligomers [25] , and by the lability of the tPA–NS complex [15] . The determination of the crystal structures of the native NS [15,35] and of its cleaved form [15] , along with recent results obtained by NMR and computer simulation [36] , have helped in recognizing some of the key dynamical and structural features of NS.…”

Section: Introductionmentioning

confidence: 99%

Functional and dysfunctional conformers of human neuroserpin characterized by optical spectroscopies and Molecular Dynamics

Noto

Santangelo

Levantino

et al. 2015

Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics

Self Cite

View full text Add to dashboard Cite

Neuroserpin (NS) is a serine protease inhibitor (SERPIN) involved in different neurological pathologies, including the Familial Encephalopathy with Neuroserpin Inclusion Bodies (FENIB), related to the aberrant polymerization of NS mutants. Here we present an in vitro and in silico characterization of native neuroserpin and its dysfunctional conformation isoforms: the proteolytically cleaved conformer, the inactive latent conformer, and the polymeric species. Based on circular dichroism and fluorescence spectroscopy, we present an experimental validation of the latent model and highlight the main structural features of the different conformers. In particular, emission spectra of aromatic residues yield distinct conformational fingerprints, that provide a novel and simple spectroscopic tool for selecting serpin conformers in vitro. Based on the structural relationship between cleaved and latent serpins, we propose a structural model for latent NS, for which an experimental crystallographic structure is lacking. Molecular Dynamics simulations suggest that NS conformational stability and flexibility arise from a spatial distribution of intramolecular salt-bridges and hydrogen bonds.

show abstract

“…Static and dynamic light scattering measurements were performed at 20 ° C by using a Brookhaven Instruments BI-9000 correlator and a solid-state laser tuned at λ 0 = 532 nm. Scattered intensity autocorrelation functions g2(t) have been analyzed by using multiple gamma functions for the diffusion coefficient and therefore by using the classic Stokes-Einstein relation to determine the size distribution P(D) of vesicles, where the size parameter D is actually the hydrodynamic radius of diffusing vesicles ( 61 ).…”

Section: Methodsmentioning

confidence: 99%

Pharmacological tuning of microRNAs in FAP-derived Extracellular Vesicles by HDAC inhibitors promotes regeneration and reduces fibrosis in dystrophic muscles

Sandonà

Consalvi

Tucciarone

et al. 2020

Preprint

View full text Add to dashboard Cite

35Functional interactions between cellular components of the muscle stem cell (MuSC) niche regulate the 36 regenerative ability of skeletal muscles in physiological and pathological conditions; however, the 37 identity of the mediators of these interactions remains largely unknown. We show here that fibro-38 adipogenic progenitor (FAP)-derived Extracellular Vesicles (EVs) mediate microRNA transfer to 39MuSCs, and that exposure of dystrophic FAPs to HDAC inhibitors (HDACi) increases the intra-EV 40 levels of a subset of microRNAs (miRs), which cooperatively target biological processes of therapeutic 41 interest, including regeneration, fibrosis and inflammation. In particular, we found that increased levels 42 of miR206 in EVs released from FAPs of muscles from Duchenne dystrophic patients or mice (mdx) 43 exposed to HDACi were associated with enhanced regeneration and inhibition of fibrosis of dystrophic 44 muscles. Consistently, EVs from HDACi-treated dystrophic FAPs could stimulated MuSC activation and 45 expansion ex vivo, and promoted regeneration, while inhibiting fibrosis and inflammation of dystrophic 46 muscles, upon intramuscular transplantation, in vivo. These data reveal a potential for pharmacological 47 modulation of FAP-derived EV's content as novel strategy for focal therapeutic interventions in 48 Duchenne Muscular Dystrophy (DMD) and possibly other muscular diseases. 49 50 51 Brief Summary 52 Extracellular Vesicles from HDACi-treated dystrophic FAPs promote regeneration, while inhibiting 53 fibrosis and inflammation of dystrophic muscles 54 55 Introduction 56Emerging evidence indicates that reciprocal interactions between distinct cellular components of 57 the regeneration machinery generate either productive or hostile environment for regeneration of 58 dystrophic muscles (1-3). While muscle stem (satellite) cells(4) (therein indicated as MuSCs) are the 59 direct effectors of muscle repair, a variety of other cell types contribute to muscle regeneration by 60 spatially and temporally coordinating the activity of MuSCs (5, 6). These cells comprise components of 61 the inflammatory infiltrate, including macrophages and eosinophils (7, 8), and a heterogeneous 62 population of muscle-derived interstitial cells referred to as fibro-adipogenic progenitors (FAPs)(9-11). 63Disruption of this network compromises the integrity of MuSC niche and has been associated with the 64 progression of many chronic muscular disorders (i.e. muscular dystrophies) and age-related decline in 65 muscle mass and repair (12, 13). 66Upon myofiber damage, FAP accumulation is preceded by the appearance of the inflammatory 67 infiltrate and is followed by MuSC activation (14, 15). This temporal pattern suggests a key role for FAPs 68 in converting inflammatory cues into signals that regulate MuSC activity, and implies reciprocal 69 communications between these cell types, through the exchange of soluble mediators, which are largely 70 unknown. Deciphering the molecular and functional identity of these mediators might reveal sel...

show abstract

The Tempered Polymerization of Human Neuroserpin

Cited by 28 publications

References 60 publications

Unlocked Concanavalin A Forms Amyloid-like Fibrils from Coagulation of Long-lived “Crinkled” Intermediates

Unlocked Concanavalin A Forms Amyloid-like Fibrils from Coagulation of Long-lived “Crinkled” Intermediates

Functional and dysfunctional conformers of human neuroserpin characterized by optical spectroscopies and Molecular Dynamics

Pharmacological tuning of microRNAs in FAP-derived Extracellular Vesicles by HDAC inhibitors promotes regeneration and reduces fibrosis in dystrophic muscles

Contact Info

Product

Resources

About