Loss of cellular homeostasis is well documented during aging and seems to be exacerbated in age-associated neurodegenerative diseases. One of the main mechanisms responsible for the maintenance of cellular homeostasis is mitophagy, which promotes the selective removal of dysfunctional mitochondria. The most known mitophagy pathway requires a coordinated set of events in which the Serine/Threonine-protein kinase PINK1, recruits the E3 ubiquitin ligase Parkin to damaged mitochondria leading to ubiquitination of outer mitochondrial membrane proteins. Ubiquitination of mitochondria acts as a signal for the recruitment of autophagy receptors required for the organelle’s degradation via autophagy. Results previously obtained in the lab, indicate that the activation of the Ca2+ -dependent phosphatase Calcineurin can act as a signal leading to Parkin recruitment to the mitochondria in the absence of PINK1. We will use a biochemical approach to validate endogenous Parkin translocation to mitochondria upon CaN activation in HEK293T cells.
In letteratura è ben documentata la perdita di omeostasi cellulare durante l’invecchiamento, ciò sembra aggravarsi con l’insorgenza di malattie neurodegenerative correlate all’invecchiamento. Uno dei principali meccanismi correlati al mantenimento dell’omeostasi cellulare è la mitofagia, essa promuove la rimozione selettiva dei mitocondri danneggiati. La pathway più conosciuta che interviene nel meccanismo di mitofagia, coinvolge una serie di eventi in cui la serina/treonina protein-chinasi PINK1 recluta l’ubiquitina ligasi E3 Parkin sulla superficie dei mitocondri danneggiati, questa ha il ruolo di ubiquitinare le proteine della membrana mitocondriale. L’ubiquitinazione dei mitocondri funge da segnale per attivare/richiamare i recettori coinvolti nella degradazione di questi organelli attraverso l’autofagia. Risultati ottenuti precedentemente in laboratorio mostrano che l’attivazione della proteina calcineurina Ca2+ fosfatasi-dipendente (CaN) può agire come segnale per il richiamo di Parkin ai mitocondri anche in assenza di PINK1. Utilizzeremo un approccio biochimico per stimolare il trasferimento di Parkin endogeno nei mitocondri tramite l’attivazione della CaN nelle cellule HEK293T.
ANALISI DELL’EFFETTO BENEFICO DELL’ATTIVAZIONE DELLA PROTEOSTASI CELLULARE IN MODELLI IN VITRO DI MALATTIA DI PARKINSON (PD)
TOSO, ADELAIDE
2022/2023
Abstract
Loss of cellular homeostasis is well documented during aging and seems to be exacerbated in age-associated neurodegenerative diseases. One of the main mechanisms responsible for the maintenance of cellular homeostasis is mitophagy, which promotes the selective removal of dysfunctional mitochondria. The most known mitophagy pathway requires a coordinated set of events in which the Serine/Threonine-protein kinase PINK1, recruits the E3 ubiquitin ligase Parkin to damaged mitochondria leading to ubiquitination of outer mitochondrial membrane proteins. Ubiquitination of mitochondria acts as a signal for the recruitment of autophagy receptors required for the organelle’s degradation via autophagy. Results previously obtained in the lab, indicate that the activation of the Ca2+ -dependent phosphatase Calcineurin can act as a signal leading to Parkin recruitment to the mitochondria in the absence of PINK1. We will use a biochemical approach to validate endogenous Parkin translocation to mitochondria upon CaN activation in HEK293T cells.File | Dimensione | Formato | |
---|---|---|---|
Toso_Adelaide.pdf
accesso riservato
Dimensione
1.51 MB
Formato
Adobe PDF
|
1.51 MB | Adobe PDF |
The text of this website © Università degli studi di Padova. Full Text are published under a non-exclusive license. Metadata are under a CC0 License
https://hdl.handle.net/20.500.12608/52046