Dissecting the differential role of C-terminal truncations in the regulation of aSyn pathology formation and the biogenesis of Lewy bodies DOI Open Access

Anne-Laure Mahul-Mellier,

Melek Firat Altay, Niran Maharjan

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 30, 2024

Alpha-synuclein (aSyn) post-translational modifications (PTMs), particularly phosphorylation at serine 129 and C-terminal truncations, are highly enriched in Lewy bodies (LBs), neurites, other types of aSyn pathological aggregates the brain patients with Parkinson's disease (PD) synucleinopathies. However, our knowledge about precise role PTMs regulating different stages pathology formation, neurodegeneration, spreading remains incomplete. In this work, we applied a systematic approach to address gap an emphasis on mapping elucidating post-fibrillization truncations uptake, processing, seeding activity, formation LB-like inclusions maturations well-established neuronal model that recapitulates all leading LB neurodegeneration. Our work shows cleavage fibrils multiple sites is conserved process occurs rapidly after during intracellular models. Interestingly, blocking internalized does not influence their whereas inhibiting enzymes regulate newly formed (e.g., calpains 1 2) significantly alters inclusions. We also show combination PTMs, play crucial interactome remodeling fibrils, including shortening, lateral association, packing maturation. Altogether, results demonstrate have differential aggregation formation. These insights, combined abundance truncated species LBs, significant implications understanding diversity developing therapeutic strategies targeting C-terminus or its proteolytic processing.

Language: Английский

Artificial intelligence–coupled plasmonic infrared sensor for detection of structural protein biomarkers in neurodegenerative diseases DOI Creative Commons
Deepthy Kavungal, Pedro Magalhães,

Senthil T. Kumar

et al.

Science Advances, Journal Year: 2023, Volume and Issue: 9(28)

Published: July 12, 2023

Diagnosis of neurodegenerative disorders (NDDs) including Parkinson's disease and Alzheimer's is challenging owing to the lack tools detect preclinical biomarkers. The misfolding proteins into oligomeric fibrillar aggregates plays an important role in development progression NDDs, thus underscoring need for structural biomarker-based diagnostics. We developed immunoassay-coupled nanoplasmonic infrared metasurface sensor that detects linked such as alpha-synuclein, with specificity differentiates distinct species using their unique absorption signatures. augmented artificial neural network enabling unprecedented quantitative prediction protein mixture. microfluidic integrated can retrieve time-resolved absorbance fingerprints presence a complex biomatrix capable multiplexing simultaneous monitoring multiple pathology-associated Thus, our promising candidate clinical diagnosis monitoring, evaluation novel therapies.

Language: Английский

Citations

50
Prominent astrocytic alpha-synuclein pathology with unique post-translational modification signatures unveiled across Lewy body disorders DOI Creative Commons
Melek Firat Altay, Alan King Lun Liu, Janice L. Holton

et al.

Acta Neuropathologica Communications, Journal Year: 2022, Volume and Issue: 10(1)

Published: Nov. 12, 2022

Abstract Alpha-synuclein (aSyn) is a pre-synaptic monomeric protein that can form aggregates in neurons Parkinson’s disease (PD), with dementia (PDD) and Lewy bodies (DLB), oligodendrocytes multiple system atrophy (MSA). Although aSyn astrocytes has previously been described PD, PDD DLB, the biochemical properties topographical distribution of astrocytic have not studied detail. Here, we present systematic investigation pathology using an expanded antibody toolset covering entire sequence key post-translational modifications (PTMs) body disorders (LBDs) MSA. Astrocytic was detected limbic cortical regions LBDs but were absent main pathological The revealed only antibodies against mid N-terminal non-amyloid component (NAC) residues 34–99. astroglial accumulations negative to canonical aggregation markers, including p62, ubiquitin pS129, positive for phosphorylated nitrated forms at Tyrosine 39 (Y39), resistant proteinase K. Our findings suggest represent major part possess distinct PTM signature characterized by both N- C-terminal truncations Y39. This first description are made solely from C-terminally cleaved species report demonstrating mixture Y39 species. These observations underscore importance characterization different cell types capture diversity brain. combined further studies on role progression pave way towards identifying novel mechanisms therapeutic targets.

Language: Английский

Citations

58
Application of Nanobiosensor Engineering in the Diagnosis of Neurodegenerative Disorders DOI Creative Commons

Thikra S. Dhahi,

Alaa Kamal Yousif Dafhalla, A. Wesam Al-Mufti

et al.

Results in Engineering, Journal Year: 2024, Volume and Issue: 24, P. 102790 - 102790

Published: Sept. 5, 2024

Language: Английский

Citations

8
A Versatile Method for Site-Specific Chemical Installation of Aromatic Posttranslational Modification Analogs into Proteins DOI Creative Commons

Xiaoxi Lin,

Shaswati Mandal, Raj V. Nithun

et al.

Published: June 17, 2024

Posttranslational modifications (PTMs) of proteins play central roles in regulating protein structure, interactome, and functions. A notable modification site is the aromatic side chain Tyr, which undergoes modifications, such as phosphorylation nitration. Despite biological physiological importance Tyr-PTMs, our current understanding mechanisms by these contribute to human health disease remains incomplete. This knowledge gap arises from absence natural amino acids that can mimic PTMs lack synthetic tools for site-specific introduction into proteins. Herein, we describe a facile method chemical installation through palladium-mediate S-C(sp2) bond formation under ambient conditions. We demonstrate incorporation novel Tyr-nitration analogs recombinantly expressed Cys-containing peptides within minutes good yield. To versatility approach, employed it prepare 10 site-specifically modified proteins, including nitrated Myc Max Furthermore, prepared focused library phosphorylated α-Syn protein, enabled first time deciphering role competing conformation aggregation vitro. Our strategy offers advantages over or semi-synthetic approaches, enables rapid selective transfer rarely explored recombinant thus facilitating generation libraries homogeneous post-translationally biomarkers discovery, mechanistic studies, drug discovery.

Language: Английский

Citations

1
Prominent astrocytic alpha-synuclein pathology with unique post-translational modification signatures unveiled across Lewy body disorders DOI Creative Commons
Melek Firat Altay, Alan King Lun Liu, Janice L. Holton

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2022, Volume and Issue: unknown

Published: May 29, 2022

ABSTRACT Alpha-synuclein (aSyn) is a pre-synaptic monomeric protein that can form aggregates in neurons Parkinson’s disease (PD), with dementia (PDD) and Lewy bodies (DLB), oligodendrocytes multiple system atrophy (MSA). Although the accumulation of aSyn astrocytes has previously been described PD, PDD DLB, biochemical properties this type pathology its topographical distribution have not studied detail. Here, we present systematic investigation astrocytic pathology, using an expanded toolset antibodies covering entire sequence known post-translational modifications (PTMs) body (LB) disorders, including sporadic PDD, familial PD SNCA G51D mutation duplication, MSA. Astrocytic was mainly detected limbic cortical regions LB but were absent key pathological These accumulations only against mid N-terminal non-amyloid component (NAC) residues 34-99. The astroglial negative to canonical aggregation markers, p62, ubiquitin pS129, positive for phosphorylated nitrated forms at Tyrosine 39 (Y39), mostly resistant proteinase K. Our findings suggest are major part possess distinct PTM signature characterized by both N- C-terminal truncations Y39. To best our knowledge, first description made solely from C-terminally cleaved species report demonstrating exists as mixture Y39 species. observations underscore critical importance characterization different cell types necessary step capturing diversity brain. combined further studies on role progression disorders pave way towards identifying novel mechanisms therapeutic targets.

Language: Английский

Citations

4
Alpha-synuclein distribution and seeding activity in rectal biopsies in Parkinson’s disease DOI Open Access
Annika Kluge,

Carmen Kintrup,

Kristína Kulcsarová

et al.

medRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 29, 2024

Abstract Background Parkinson’s disease (PD) is characterized by the accumulation of alpha-synuclein (aSyn) pathology, not only in brain but also gastrointestinal (GI) tract. This study investigates use unique aSyn antibodies and an seed amplification assay (SAA) for detecting pathological rectal biopsy samples from PD patients healthy individuals. These were preserved using formalin-fixed paraffin-embedded (FFPE) methods. Materials Methods The analyzed seeding capacity FFPE submucosal biopsies 24 20 controls aSyn-SAA. distribution was examined immunohistochemistry with targeting specific conformations phosphorylated forms at S129 Y39. Results Pathological found all patients, as confirmed SAA, these linked to severity motor symptoms (MDS-UPDRS-III). However, immunoreactive patterns conformation-specific or did show notable differences between subjects. Conclusion strains are detectable high accuracy utility immunohistochemical detection current identifying appears limited. findings advocate aSyn-SAA a diagnostic tool PD, contributing deeper understanding gut-brain connection disease.

Language: Английский

Citations

0
Dissecting the differential role of C-terminal truncations in the regulation of aSyn pathology formation and the biogenesis of Lewy bodies DOI Open Access

Anne-Laure Mahul-Mellier,

Melek Firat Altay, Niran Maharjan

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 30, 2024

Alpha-synuclein (aSyn) post-translational modifications (PTMs), particularly phosphorylation at serine 129 and C-terminal truncations, are highly enriched in Lewy bodies (LBs), neurites, other types of aSyn pathological aggregates the brain patients with Parkinson's disease (PD) synucleinopathies. However, our knowledge about precise role PTMs regulating different stages pathology formation, neurodegeneration, spreading remains incomplete. In this work, we applied a systematic approach to address gap an emphasis on mapping elucidating post-fibrillization truncations uptake, processing, seeding activity, formation LB-like inclusions maturations well-established neuronal model that recapitulates all leading LB neurodegeneration. Our work shows cleavage fibrils multiple sites is conserved process occurs rapidly after during intracellular models. Interestingly, blocking internalized does not influence their whereas inhibiting enzymes regulate newly formed (e.g., calpains 1 2) significantly alters inclusions. We also show combination PTMs, play crucial interactome remodeling fibrils, including shortening, lateral association, packing maturation. Altogether, results demonstrate have differential aggregation formation. These insights, combined abundance truncated species LBs, significant implications understanding diversity developing therapeutic strategies targeting C-terminus or its proteolytic processing.

Language: Английский

Citations

0