PXD061021

PXD061021 is an original dataset announced via ProteomeXchange.

Title	Molecular Structure and Enzymatic Mechanism of the Human Collagen Hydroxylysine Galactosyltransferase GLT25D1/COLGALT1
Description	During collagen biosynthesis, lysine residues undergo extensive post-translational modifications through the alternate action of two distinct metal ion-dependent enzyme families (i.e., LH/PLODs and GLT25D/COLGALT), ultimately producing the highly conserved alpha-(1,2)-glucosyl-beta-(1,2)-galactosyl-5-hydroxylysine pattern. Malfunctions in these enzymes are linked to developmental pathologies and extracellular matrix alterations associated to enhanced aggressiveness of solid tumors. Here, we characterized human GLT25D1/COLGALT1, revealing an elongated head-to-head homodimeric assembly. Each monomer encompasses two domains (GT1 and GT2), both unexpectedly capable of binding metal ion cofactors and UDP-alpha-galactose donor substrates, resulting in four candidate catalytic sites per dimer. We identified the catalytic site in GT2, featuring an unusual Glu-Asp-Asp motif critical for Mn2+ binding, ruling out direct catalytic roles for the GT1 domain, but showing that the unexpectedly bound Ca2+ and UDP-alpha-galactose cofactors are critical for folding stability. Dimerization was not essential for GLT25D1/COLGALT1 activity, but rather a hallmark for multi-enzyme assembly interactions and/or collagen substrate recognition.
HostingRepository	PRIDE
AnnounceDate	2025-03-05
AnnouncementXML	Submission_2025-03-05_02:17:40.921.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD061021
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alice Vetrano
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-20 14:09:01	ID requested
⏵ 1	2025-03-05 02:17:41	announced

10.6019/PXD061021;

submitter keyword: Glycosyltransferase,Collagen Biosynthesis

Extracellular Matrix

Post-translational modifications

Claudio Iacobucci
contact affiliation	University of L'Aquila
contact email	claudio.iacobucci@univaq.it
lab head
Alice Vetrano
contact affiliation	University of L'Aquila
contact email	alice.vetrano@univaq.it
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/03/PXD061021

PRIDE project URI

• PRIDE
  1. PXD061021
     1. Label: PRIDE project
     2. Name: Molecular Structure and Enzymatic Mechanism of the Human Collagen Hydroxylysine Galactosyltransferase GLT25D1/COLGALT1