PXD064332-1

PXD064332 is an original dataset announced via ProteomeXchange.

Title	Small ORF16 modulates the Mtr complex activity in Methanosarcina mazei depending on the molecular hydrogen availability
Description	Small open reading frame (small ORF)-encoded proteins, with less than 100 amino acids in length, have attracted increasing attention over the past decade after being largely overlooked due to limitations in classical bioinformatics and biochemical methodologies. For the mesophilic archaeal model system Methanosarcina mazei, a high number of previously unannotated small ORFs have been identified through ribosome profiling combined with LC-MS/MS analysis. However, the physiological role of the majority of the respective small proteins remains unknown. Here, we report on the functional characterization of the small ORF16 encoded small protein MtrR (49 amino acids). We demonstrated that MtrR is forming oligomers localized at the cytoplasmic membrane. There, it interacts with the tetrahydrosarcinapterin S-methyltransferase (Mtr), a key membrane-bound complex of the energy metabolism, and impacts its activity. In vitro interaction and in vivo copurification assays revealed an interaction of MtrR with the Mtr-complex, which was further validated by microscale thermophoresis analysis demonstrating a specific strong interaction with the MtrA subunit. Analyzing growth under varying molecular hydrogen (H2) availability demonstrated that the small ORF16 deletion mutant showed significantly impaired growth in the presence of H2, independent of the carbon source. Consequently, we propose that MtrR fine-tunes the activity of the Mtr-complex in response to fluctuating H2 availabilities, allowing to adapt the energy metabolism to changing environmental H2 conditions.
HostingRepository	PRIDE
AnnounceDate	2026-09-07
AnnouncementXML	Submission_2026-09-06_16:21:53.906.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andreas Tholey
SpeciesList	scientific name: Methanosarcina mazei Go1; NCBI TaxID: NEWT:192952;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2025-05-27 06:35:15	ID requested
⏵ 1	2026-09-06 16:21:54	announced

10.1111/febs.70457;

Habenicht T, Hastedt B, Cassidy L, Kie, ß, ling C, Tholey A, Schuller JM, Schmitz RA, -dependent modulation of tetrahydromethanopterin S-methyltransferase (Mtr complex) activity by the small protein MtrR in Methanosarcina mazei. FEBS J, 293(14):4397-4410(2026) [pubmed]

submitter keyword: tetrahydrosarcinapterin S-methyltransferase (Mtr), small ORF encoded proteins,Methanosarcina mazei, LC MSMS

Prof. Dr. Andreas Tholey
contact affiliation	AG Proteomics & Bioanalytics - Institut für Experimentelle Medizin Christian-Albrechts-Universität zu Kiel 24105 Kiel, Germany
contact email	A.Tholey@iem.uni-kiel.de
lab head
Andreas Tholey
contact affiliation	Systematic Proteome Research & Bioanalytics, University of Kiel
contact email	a.tholey@iem.uni-kiel.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD064332
     1. Label: PRIDE project
     2. Name: Small ORF16 modulates the Mtr complex activity in Methanosarcina mazei depending on the molecular hydrogen availability