PXD026486-1

PXD026486 is an original dataset announced via ProteomeXchange.

Title	Structural and functional fine mapping of cysteines in mammalian glutaredoxin reveal a hierarchy of susceptibility to oxidative inactivation
Description	Protein-S-glutathionylation (PSSG) is a post-translational modification involving the conjugation of glutathione to reactive protein thiols, which can modulate the activity and structure of key cellular proteins. Glutaredoxins (GLRX) are oxidoreductases that regulate this process by performing the reversible deglutathionylation reaction. Mammalian GLRX only requires the N-terminal cysteine in the thioredoxin domain to carry out deglutathionylation. However, GLRX has five cysteines that are potentially vulnerable to oxidative modification, and oxidation of GLRX itself has been reported in close association with aggregation and loss of activity. Further, the specific cysteines within GLRX that are vulnerable to oxidative modification and their relative reactivities with oxidants remain unknown. Herein, we utilized various molecular modeling approaches coupled with site-directed mutagenesis of each cysteine both individually and in combination to address how the five cysteines present in GLRX influence the enzyme’s biological activity and susceptibility to dimerization and inactivation. We report that mutation of cysteine 26 (C26) to serine of murine GLRX1 confers a higher rate of deglutathionylation. We further demonstrate that C8 and C83 are targets for PSSG in vitro while C79 is less vulnerable due to steric hindrance. Hydrogen peroxide also caused inactivation of GLRX that was only protected against with mutation of C8 or C83. Both experimental and modeling evidence indicates C8 contributes to dimer formation. Lastly, combinatorial mutation of C8, C26, and C83 results in increased biological activity and resistance to aggregation and oxidative inactivation. Overall, these results from our corroborated experimental and computational studies have valuable implications for the use of GLRX as a therapeutic in settings of dysregulated protein glutathionylation.
HostingRepository	PRIDE
AnnounceDate	2023-10-24
AnnouncementXML	Submission_2023-10-24_11:13:29.990.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD026486
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Ying WaiLam
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-06-04 03:02:18	ID requested
⏵ 1	2023-10-24 11:13:30	announced
2	2023-11-14 09:07:24	announced	2023-11-14: Updated project metadata.
3	2024-10-22 06:08:43	announced	2024-10-22: Updated project metadata.

Corteselli EM, Sharafi M, Hondal R, MacPherson M, White S, Lam YW, Gold C, Manuel AM, van der Vliet A, Schneebeli ST, Anathy V, Li J, Janssen-Heininger YMW, Structural and functional fine mapping of cysteines in mammalian glutaredoxin reveal their differential oxidation susceptibility. Nat Commun, 14(1):4550(2023) [pubmed]

10.6019/PXD026486;

submitter keyword: molecular dynamics, glutathione, multiscale modeling, redox,S-glutathionylation, quantum mechanics

YvonneJanssen-Heininger
contact affiliation	Department of Pathology and Laboratory Medicine University of Vermont Larner College of Medicine Burlington, Vermont, 05405, USA
contact email	yvonne.janssen@med.uvm.edu
lab head
Ying WaiLam
contact affiliation	Vermont Genetics Network Proteomics Facility Department of Biology University of Vermont
contact email	ylam@uvm.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD026486
     1. Label: PRIDE project
     2. Name: Structural and functional fine mapping of cysteines in mammalian glutaredoxin reveal a hierarchy of susceptibility to oxidative inactivation