Fluorescence Methods Applied to the Description of Urea-Dependent YME1L Protease Unfolding.
AAA+ protease
ATP-dependent protease
YME1
mitochondria
protein unfolding
proteostasis
Journal
Biomolecules
ISSN: 2218-273X
Titre abrégé: Biomolecules
Pays: Switzerland
ID NLM: 101596414
Informations de publication
Date de publication:
23 04 2020
23 04 2020
Historique:
received:
20
02
2020
revised:
20
04
2020
accepted:
21
04
2020
entrez:
29
4
2020
pubmed:
29
4
2020
medline:
7
4
2021
Statut:
epublish
Résumé
ATP-dependent proteases are ubiquitous across all kingdoms of life and are critical to the maintenance of intracellular protein quality control. The enzymatic function of these enzymes requires structural stability under conditions that may drive instability and/or loss of function in potential protein substrates. Thus, these molecular machines must demonstrate greater stability than their substrates in order to ensure continued function in essential quality control networks. We report here a role for ATP in the stabilization of the inner membrane YME1L protease. Qualitative fluorescence data derived from protein unfolding experiments with urea reveal non-standard protein unfolding behavior that is dependent on [ATP]. Using multiple fluorophore systems, stopped-flow fluorescence experiments demonstrate a depletion of the native YME1L ensemble by urea-dependent unfolding and formation of a non-native conformation. Additional stopped-flow fluorescence experiments based on nucleotide binding and unfoldase activities predict that unfolding yields significant loss of active YME1L hexamers from the starting ensemble. Taken together, these data clearly define the stress limits of an important mitochondrial protease.
Identifiants
pubmed: 32340357
pii: biom10040656
doi: 10.3390/biom10040656
pmc: PMC7226517
pii:
doi:
Substances chimiques
Anilino Naphthalenesulfonates
0
Nucleotides
0
Solvents
0
1-anilino-8-naphthalenesulfonate
630I4V6051
Tryptophan
8DUH1N11BX
Urea
8W8T17847W
Peptide Hydrolases
EC 3.4.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Déclaration de conflit d'intérêts
Linear Extrapolation Method
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