A5084150671- Genetic Neurodegenerative Diseases
- Enzyme Structure and Function
- Alzheimer's disease research and treatments
- Mitochondrial Function and Pathology
- Biochemical and Molecular Research
- Amyloidosis: Diagnosis, Treatment, Outcomes
- Protein Structure and Dynamics
- Monoclonal and Polyclonal Antibodies Research
- RNA modifications and cancer
- Signaling Pathways in Disease
- Glycosylation and Glycoproteins Research
- Fungal and yeast genetics research
- RNA and protein synthesis mechanisms
- Advanced Electron Microscopy Techniques and Applications
- Cellular transport and secretion
- Force Microscopy Techniques and Applications
- Tea Polyphenols and Effects
- RNA regulation and disease
- Prion Diseases and Protein Misfolding
- Cancer-related gene regulation
- vaccines and immunoinformatics approaches
- Ubiquitin and proteasome pathways
- Microbial Metabolic Engineering and Bioproduction
- Escherichia coli research studies
- Nuclear Structure and Function
University of Milan
2019-2025
IRCCS Policlinico San Donato
2022-2023
University of California, Berkeley
2022
University of Milano-Bicocca
2015-2018
Universitat Autònoma de Barcelona
2018
Institut de Biosciences et Biotechnologies
2014
Abstract Monoclonal antibodies have emerged as key therapeutics. In particular, nanobodies, small, single-domain that are naturally expressed in camelids, rapidly gaining momentum following the approval of first nanobody drug 2019. Nonetheless, development these biologics therapeutics remains a challenge. Despite availability established vitro directed-evolution technologies relatively fast and cheap to deploy, gold standard for generating therapeutic discovery from animal immunization or...
Abstract hnRNPDL is a ribonucleoprotein (RNP) involved in transcription and RNA-processing that hosts missense mutations causing limb-girdle muscular dystrophy D3 (LGMD D3). Mammalian-specific alternative splicing (AS) renders three natural isoforms, hnRNPDL-2 being predominant humans. We present the cryo-electron microscopy structure of full-length amyloid fibrils, which are stable, non-toxic, bind nucleic acids. The high-resolution core consists single Gly/Tyr-rich highly hydrophilic...
The polyglutamine (polyQ)-containing protein ataxin-3 (AT3) triggers the neurodegenerative disease spinocerebellar ataxia type 3 (SCA3) when its polyQ tract is expanded beyond a critical length. This results in aggregation and generation of toxic oligomers fibrils. Currently, no effective treatment available for such other diseases. Therefore, plenty investigations are being carried on to assess mechanism action therapeutic potential anti-amyloid agents. polyphenol compound...
Accurate measurements of binding kinetics, encompassing equilibrium dissociation constant (KD), association rate (kon), and (koff), are critical for the development optimisation high-affinity proteins. However, such require highly purified material precise ligand immobilisation, limiting number binders that can be characterised within a reasonable timescale budget. Here, we present SpyBLI method, rapid cost-effective biolayer interferometry (BLI) pipeline leverages SpyCatcher003-SpyTag003...
Protein aggregation into amyloid fibrils is the archetype of aberrant biomolecular self-assembly processes, with more than 50 associated diseases that are mostly uncurable. Understanding mechanisms thus fundamental importance and goes in parallel structural characterization transient oligomers formed during process. Oligomers have been proven elusive to high-resolution techniques, while large sizes long time scales, typical limited use computational methods date. To surmount these...
The protein ataxin-3 (ATX3) triggers an amyloid-related neurodegenerative disease when its polyglutamine stretch is expanded beyond a critical threshold. We formerly demonstrated that the polyphenol epigallocatechin-3-gallate (EGCG) could redirect amyloid aggregation of full-length, ATX3 (ATX3-Q55) towards non-toxic, soluble, SDS-resistant aggregates. Here, we have characterized other related phenol compounds, although smaller in size, i.e. (-)-epigallocatechin gallate (EGC), and gallic acid...
Abstract Epigallocatechin‐3‐gallate (EGCG) and tetracycline are two known inhibitors of amyloid aggregation able to counteract the fibrillation most proteins involved in neurodegenerative diseases. We have recently investigated their effect on ataxin‐3 (AT3), polyglutamine‐containing protein responsible for spinocerebellar ataxia type 3. previously showed that EGCG can contrast process toxicity expanded AT3, although by different mechanisms. Here, we performed further experiments using sole...
Abstract Monoclonal antibodies have emerged as key therapeutics, and nanobodies are rapidly gaining momentum following the approval of first nanobody drug in 2019. Nonetheless, development these biologics therapeutics remains a challenge. Despite availability established vitro directed evolution technologies that relatively fast cheap to deploy, gold standard for generating therapeutic discovery from animal immunization or patients. Immune-system derived tend favourable properties vivo,...
The molecular bases of amyloid aggregation propensity are still poorly understood, especially for proteins that display a stable folded native structure. A prototypic example is human beta-2 microglobulin (β2m), which, when accumulated in patients, gives rise to dialysis-related amyloidosis. Interestingly, although the physiologic concentration β2m mice five times higher than found no deposits observed mice. Moreover, murine (mβ2m) not only displays lower both vivo and vitro but also...
Familial encephalopathy with neuroserpin inclusion bodies (FENIB) is a severe and lethal neurodegenerative disease. Upon specific point mutations in the SERPINI1gene-coding for human protein (NS) resulting pathologic NS variants polymerize accumulate within endoplasmic reticulum of neurons central nervous system. To date, embelin (EMB) only known inhibitor polymerization vitro. This molecule capable preventing dissolving preformed polymers. Here, we show that lowering EMB concentration...
Ataxin-3 (AT3) is a deubiquitinating enzyme that triggers an inherited neurodegenerative disorder, spinocerebellar ataxia type 3, when its polyglutamine (polyQ) stretch close to the C-terminus exceeds critical length. AT3 variants carrying expanded polyQ are prone associate with each other into amyloid toxic aggregates, which responsible for neuronal death ensuing neurodegeneration. We employed Saccharomyces cerevisiae as eukaryotic cellular model better clarify mechanism by disease....
Neuroserpin (NS) is a member of the serine protease inhibitors superfamily. Specific point mutations are responsible for its accumulation in endoplasmic reticulum neurons that leads to pathological condition named familial encephalopathy with neuroserpin inclusion bodies (FENIB). Wild-type NS presents two N-glycosylation chains and does not form polymers vivo, while non-glycosylated causes aberrant polymer cell models. To date, all vitro studies have been conducted on bacterially expressed...
Genetically-encoded combinatorial peptide libraries are convenient tools to identify peptides be used as therapeutics, antimicrobials and functional synthetic biology modules. Here, we report the identification characterization of a cyclic peptide, G4CP2, that interferes with GAL4 protein, transcription factor responsible for activation galactose catabolism in yeast widely exploited molecular biology. G4CP2 was identified by screening CYCLIC, Yeast Two-Hybrid-based library developed our...
Abstract Reactive Intermediate Deaminase (Rid) protein superfamily includes eight families among which the RidA is conserved in all domains of life. proteins accelerate deamination reactive 2-aminoacrylate (2AA), an enamine produced by some pyridoxal phosphate (PLP)-dependent enzymes. 2AA accumulation inhibits target enzymes with a detrimental impact on fitness. As consequence whole genome duplication, teleost fish have two ridA paralogs, while other extant vertebrates contain single-copy...
Amyloid aggregation of human ataxin-3 (ATX3) is responsible for spinocerebellar ataxia type 3, which belongs to the class polyglutamine neurodegenerative disorders. It widely accepted that formation toxic oligomeric species primarily involved in onset disease. For this reason, understand mechanisms underlying toxicity, we expressed both a physiological (ATX3-Q24) and pathological ATX3 variant (ATX3-Q55) simplified cellular model, Escherichia coli. has been observed ATX3-Q55 expression...
The protein ataxin-3 contains a polyglutamine stretch that triggers amyloid aggregation when it is expanded beyond critical threshold. This results in the onset of spinocerebellar ataxia type 3. consists globular N-terminal Josephin domain and disordered C-terminal tail where located. Expanded aggregates via two-stage mechanism: first, self-association, then polyQ fibrillation. highlights intrinsic amyloidogenic potential domain. Therefore, much effort has been put into investigating its...
Reactive intermediate deaminase A (RidA) is a highly conserved enzyme that catalyzes the hydrolysis of 2-imino acids to corresponding 2-keto and ammonia. RidA thus prevents accumulation such potentially harmful compounds in cell, as exemplified by its role degradation 2-aminoacrylate, formed during metabolism cysteine serine, catalyzing conversion stable 2-iminopyruvate tautomer into pyruvate. Capra hircus (goat) (
Human PNPase (hPNPase) is an essential RNA exonuclease located in mitochondria, where it contributes to import from the cytoplasm, degradation of mitochondrial RNA, and R-loop homeostasis. Biallelic mutations hPNPase PNPT1 gene cause different genetic diseases, ranging hereditary hearing loss Leigh syndrome. In this work, we used Escherichia coli model recently developed test effects four pathological associated with diseases severity. Moreover, generated a new human cell by introducing into...
Abstract Human PNPase (hPNPase) is an essential RNA exonuclease located in mitochondria, where it contributes to import from the cytoplasm, degradation of mitochondrial and R-loop homeostasis. Biallelic mutations hPNPase PNPT1 gene cause different genetic diseases, ranging hereditary hearing loss Leigh syndrome. In this work, we used Escherichia coli model test effects four pathological associated with diseases severity. Moreover, generated a new human cell by introducing into 293T cells via...
Abstract Protein aggregation into amyloid fibrils is the archetype of aberrant biomolecular self-assembly processes, with more than 50 diseases associated that are mostly uncurable. Understanding mechanisms thus fundamental importance and goes in parallel characterization structures transient oligomers formed process. Oligomers have been proven elusive to high-resolution structural techniques, while large sizes long-time scales typical processes limited, so far, use computational methods. To...