PXD053751

PXD053751 is an original dataset announced via ProteomeXchange.

Title	Site-specific quantification of collagen post-translational modifications reveals profound disruption of the collagen biosynthetic network upon prolyl-3-hydroxylase 1 deficiency
Description	Type I collagen is the main structural protein in mammals and other vertebrate life forms. During the biosynthesis of collagens, prolyl-3-hydroxylase 1 (P3H1) catalyzes 3-hydroxylation of proline residues and acts as a chaperone. Deficiency of P3H1 causes Osteogenesis Imperfecta (OI), a hereditary disorder primarily associated with mutations in type I collagen and collagen-related genes. However, the full impact of P3H1 deficiency on the collagen biosynthesis machinery and post-translational modifications (PTMs) is not known. In this comprehensive study, we conducted a detailed investigation of P3h1 knockout mice collagen by performing amino acid analysis, high-resolution tandem mass spectrometry (MS/MS)-based identification and site-specific quantification of collagen PTMs, and gene expression analysis to unravel the impact of P3H1 deficiency on type I collagen biosynthesis in mouse tail tendon. Amino acid analysis revealed a general overmodification of prolines and lysines in mouse tendon type I collagen. Site-specific MS/MS-based quantification of collagen PTMs verified the P3H1-dependent 3-hydroxyproline sites COL1A1-P1153 and COL1A2-P803, but demonstrated frequent overhydroxylation of prolines, including 3- and 4-hydroxylation, as well as increased 5-hydroxylation of lysines in unprecedented detail. Gene expression analysis in P3H1-deficient mouse tail tenocytes unveiled upregulation of Col1a1 expression along with several other genes encoding collagen biosynthetic enzymes. Interestingly, upregulation of P4ha2 expression stood out and correlated with specific overhydroxylation of prolyl-4-hydroxylation sites containing the P4HA2 preferred motif EPG. In summary, the loss of P3H1 led to a profound imbalance in the entire ER-resident collagen biosynthetic machinery and drastically altered the collagen I PTM network. In conclusion, our findings offer a novel mechanism underlying overmodification in addition to prolonged exposure of unfolded collagen chains in the ER, suggest the presence of so far unknown feedback mechanisms between ER-resident collagen biosynthesis and collagen biosynthetic gene expression, and further emphasize the essential role of P3H1 in maintaining collagen quality control.
HostingRepository	PRIDE
AnnounceDate	2026-04-02
AnnouncementXML	Submission_2026-04-02_11:06:58.486.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Claudia Staab-Weijnitz
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: NEWT:10090;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2024-07-08 04:40:57	ID requested
⏵ 1	2026-04-02 11:06:59	announced

10.1016/J.JBC.2026.111422;

submitter keyword: glycosylation,collagen, tendon, post-translational modification (PTM), prolyl-3-hydroxylase, hydroxyproline

Claudia A.
contact affiliation	Institute of Lung Health and Immunity (LHI), Helmholtz Munich, Comprehensive Pneumology Center (CPC-M), Germany, Member of the German Center for Lung Research (DZL); Department of Pediatrics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA
contact email	claudia.staab-weijnitz@cuanschutz.edu
lab head
Claudia Staab-Weijnitz
contact affiliation	University of Colorado, Anschutz Medical Campus
contact email	claudia.staab-weijnitz@cuanschutz.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD053751
     1. Label: PRIDE project
     2. Name: Site-specific quantification of collagen post-translational modifications reveals profound disruption of the collagen biosynthetic network upon prolyl-3-hydroxylase 1 deficiency