Phosphorylation of Prion Protein at Serine 43 Induces Prion Protein Conformational Change

Giannopoulos, Paresa N.; Robertson, Catherine; Jodoin, Julie; Paudel, Hemant K.; Booth, Stephanie A.; LeBlanc, Andréa C.

doi:10.1523/jneurosci.2294-09.2009

Cited by 25 publications

(24 citation statements)

References 53 publications

(63 reference statements)

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“…Therefore we suggest an aggregation scheme (depicted in and outlined the host of far-reaching effects, either on molecular or on cellular levels. [19][20][21]25,[67][68][69] Our results demonstrated that Trp-cage miniproteins are sensitive models for studying the effect of phosphorylation on aggregation and amyloid formation, exhibiting highly site-specific effects, in some cases facilitating and in others by blocking amyloid fibrillization. (Table S1), Sequence analysis for amylidogenic regions of Trpcage (Fig.…”

Section: Ph Dependence Of the Aggregation Of D9s(p) And 13s(p) Trp-camentioning

confidence: 69%

Phosphorylation as Conformational Switch from the Native to Amyloid State: Trp-Cage as a Protein Aggregation Model

et al. 2015

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The 20 residue long Trp-cage miniprotein is an excellent model both for computational and experimental studies of protein folding and stability. Recently, a great attention emerged to study disease-related protein misfolding, aggregation and amyloid formation, with the aim of revealing their structural and thermodynamic background. Trp-cage is sensitive to both environmental and structure-modifying effects, and aggregates with ease upon structure destabilization and thus, it is an ideal model of aggregation and amyloid formation. Here, we characterize the amyloid formation of several sequence modified and side-chain phosphorylated Trp-cage variants. We applied NMR, CD, FTIR spectroscopy, MD simulations, transmission electron microscopy in conjunction with thioflavin-T fluorescence measurements to reveal the structural consequences of side-chain phosphorylation. We demonstrate that the native fold is destabilized upon serine phosphorylation and the resultant highly dynamic structures form amyloid-like ordered aggregates with high intermolecular β-structure content. The only exception is the D9S(P) variant which follows an alternative aggregation process by forming thin fibrils, presenting a CD spectrum of PPII helix, and showing low ThT binding capability. We propose a complex aggregation model for these Trp-cage miniproteins. This model assumes an additional aggregated state, a collagen triple helical form which can precede amyloid formation. The phosphorylation of a single serine residue serves as a conformational switch, triggering aggregation, otherwise mediated by kinases in cell. We show that Trp-cage miniprotein is indeed a realistic model of larger globular systems of composite folding and aggregation landscapes and helps us to understand the fundamentals of deleterious protein aggregation and amyloid formation.3

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Section: Ph Dependence Of the Aggregation Of D9s(p) And 13s(p) Trp-camentioning

confidence: 69%

Phosphorylation as Conformational Switch from the Native to Amyloid State: Trp-Cage as a Protein Aggregation Model

et al. 2015

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“…Reaction-Previously, we have shown that the Cdk5-phosphorylated PrP was proteinase K-resistant and Congo Red-positive, and transmission electron microscopy revealed that Cdk5-phosphorylated PrP WT converted into aggregates composed of both anti-pPrP S43 immunopositive and immunonegative globules, which became compacted and generated rare fibrils with aging (15). The hyperbolic shape of the IKTA reaction with full-length wild type phosphorylated PrPs indicates a rapid conversion of these PrPs after Cdk5 phosphorylation (Fig.…”

Section: The Cdk5-phosphorylated Prp Wt-v Conversion Is Reversed Uponmentioning

confidence: 87%

“…The phosphorylated PrPs may convert irreversibly with time. We have shown by electron microscopy that aging of the phosphorylated wild type PrP resulted in a change from globular aggregates to a more compact structure (15). The increased rate constant of Cdk5-mediated PrP conversion in the familial PrP mutants may explain the age-dependent manifestation of the prion diseases associated with those mutations.…”

Section: Discussionmentioning

confidence: 99%

“…The prokaryotically expressed purified recombinant C-terminally His-tagged mutant PrPs were over 98% pure as observed by Coomassie Blue-stained SDS-PAGE. As described previously (15), purified wild type PrP with a valine codon 129 (PrP WT-V ) was phosphorylated by Cdk5, whereas mutant PrP…”

Section: Familial Prp Mutants Are Equally and Efficientlymentioning

confidence: 99%

“…PrP is phosphorylated by various kinases, including Fyn and casein kinase II (14). PrP is also phosphorylated at Ser 43 (pPrP S43 ) by neuronal cyclin-dependent kinase 5 (Cdk5), and pPrP S43 forms proteinase K-resistant aggregates and accumulates in scrapie-infected mouse brains (15). Cdk5 levels are increased in PrP SC -infected brains (16), and the p25/p35 activators of Cdk5 and Cdk5 increase with age (17).…”

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confidence: 99%

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Cyclin-dependent Kinase 5 Phosphorylation of Familial Prion Protein Mutants Exacerbates Conversion into Amyloid Structure

Rouget

Sharma

LeBlanc

2015

Journal of Biological Chemistry

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Background: Conversion of prion protein occurs in familial prion diseases, but the exact mechanism is unknown. Results: Phosphorylation at the N-terminal serine 43 causes or exacerbates conversion of familial prion protein mutants. Conclusion:The N terminus and C terminus of prion protein both influence conversion into amyloid. Significance: The flexible N terminus of prion protein plays a role in amyloid conversion and could indicate a novel target to prevent prion conversion.

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Big versus small: The impact of aggregate size in disease

et al. 2023

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Protein aggregation results in an array of different size soluble oligomers and larger insoluble fibrils. Insoluble fibrils were originally thought to cause neuronal cell deaths in neurodegenerative diseases due to their prevalence in tissue samples and disease models. Despite recent studies demonstrating the toxicity associated with soluble oligomers, many therapeutic strategies still focus on fibrils or consider all types of aggregates as one group. Oligomers and fibrils require different modeling and therapeutic strategies, targeting the toxic species is crucial for successful study and therapeutic development. Here, we review the role of different‐size aggregates in disease, and how factors contributing to aggregation (mutations, metals, post‐translational modifications, and lipid interactions) may promote oligomers opposed to fibrils. We review two different computational modeling strategies (molecular dynamics and kinetic modeling) and how they are used to model both oligomers and fibrils. Finally, we outline the current therapeutic strategies targeting aggregating proteins and their strengths and weaknesses for targeting oligomers versus fibrils. Altogether, we aim to highlight the importance of distinguishing the difference between oligomers and fibrils and determining which species is toxic when modeling and creating therapeutics for protein aggregation in disease.

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Phosphorylation of Prion Protein at Serine 43 Induces Prion Protein Conformational Change

Cited by 25 publications

References 53 publications

Phosphorylation as Conformational Switch from the Native to Amyloid State: Trp-Cage as a Protein Aggregation Model

Phosphorylation as Conformational Switch from the Native to Amyloid State: Trp-Cage as a Protein Aggregation Model

Cyclin-dependent Kinase 5 Phosphorylation of Familial Prion Protein Mutants Exacerbates Conversion into Amyloid Structure

Big versus small: The impact of aggregate size in disease

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