COLGALT1

UniProt ID: Q8NBJ5
Organism: Homo sapiens
Review Status: COMPLETE
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Gene Description

COLGALT1 (collagen beta(1-O)galactosyltransferase 1; also known as GLT25D1) is a soluble enzyme of the endoplasmic reticulum lumen that catalyzes the first committed step of collagen O-linked glycosylation. It transfers galactose from UDP-alpha-D-galactose onto (5R)-5-hydroxy-L-lysine (hydroxylysine) residues of collagen, forming the beta(1-O)-linked Gal-O-hydroxylysine (EC 2.4.1.50). This galactose is then the acceptor for subsequent alpha1,2-glucosylation, generating the Glc(alpha1-2)Gal disaccharide that is strongly conserved across animal collagens. The enzyme belongs to glycosyltransferase family 25 (GT25), adopts a GT-A-like fold with metal-dependent (Mn2+) catalysis, and uses essential aspartate residues (D166/D168, D461/D463) for activity. COLGALT1 acts on multiple collagen types (including types I and IV) as well as the collagenous domain of mannose-binding lectin, and co-localizes in the early secretory pathway with the upstream lysyl hydroxylase PLOD3/LH3. As the predominant of two paralogous collagen galactosyltransferases (the other being COLGALT2/GLT25D2), it is broadly expressed and its loss reduces collagen glycosylation, causing intracellular collagen accumulation. Biallelic loss-of-function variants in COLGALT1 cause an autosomal recessive cerebral small vessel disease (brain small vessel disease 3).

Existing Annotations Review

GO Term Evidence Action Reason
GO:0050211 procollagen galactosyltransferase activity
IBA
GO_REF:0000033
ACCEPT
Summary: Phylogenetically inferred procollagen galactosyltransferase activity. This is the well-established core molecular function of COLGALT1, directly demonstrated experimentally for the human enzyme.
Reason: This is the core molecular function of COLGALT1, supported by direct biochemical characterization (EC 2.4.1.50) and mutagenesis. The IBA transfer is fully consistent with the experimental evidence.
Supporting Evidence:
PMID:19075007
Amino acid analysis of the products of GLT25D1 and GLT25D2 reactions confirmed the transfer of galactose to hydroxylysine residues.
GO:0005788 endoplasmic reticulum lumen
IEA
GO_REF:0000044
ACCEPT
Summary: ER lumen localization mapped from UniProt subcellular location. COLGALT1 is a soluble ER-lumen protein, directly shown by immunofluorescence and membrane-floatation assays.
Reason: ER lumen is the correct site of action; this electronic mapping agrees with direct experimental localization evidence.
Supporting Evidence:
PMID:20470363
In agreement with the predictions our results show that GLT25D1 is directed to the ER lumen as a soluble protein and retained there.
GO:0050211 procollagen galactosyltransferase activity
IEA
GO_REF:0000120
ACCEPT
Summary: Electronically inferred procollagen galactosyltransferase activity (mapped from RHEA:12637 / EC 2.4.1.50). This is the experimentally validated core function.
Reason: Consistent with the experimentally demonstrated catalytic activity and EC assignment.
Supporting Evidence:
file:human/COLGALT1/COLGALT1-uniprot.txt
EC=2.4.1.50; Reaction=(5R)-5-hydroxy-L-lysyl-[collagen] + UDP-alpha-D-galactose = (5R)-5-O-(beta-D-galactosyl)-5-hydroxy-L-lysyl-[collagen] + UDP + H(+); Xref=Rhea:RHEA:12637.
GO:0030199 collagen fibril organization
TAS
Reactome:R-HSA-1650814
KEEP AS NON CORE
Summary: COLGALT1 contributes to collagen biosynthesis/modification, which is upstream of collagen fibril organization in the extracellular matrix. Its direct action is intracellular hydroxylysine galactosylation, not fibril assembly itself.
Reason: This is a downstream, indirect biological consequence of collagen glycosylation rather than the core molecular function. Glycosylation has been suggested to influence collagen crosslink formation and matrix organization, but the enzyme does not directly organize fibrils. Retaining as non-core captures the pathway context without overstating the role.
Supporting Evidence:
PMID:27402836
glycosylation may be involved in the organization of collagens in the extracellular space. Glycosylation has been suggested to regulate cross-link formation in fibrillar collagens.
GO:0050211 procollagen galactosyltransferase activity
TAS
Reactome:R-HSA-1981120
ACCEPT
Summary: Reactome-asserted procollagen galactosyltransferase activity, the core molecular function of COLGALT1.
Reason: Core molecular function, well supported by direct biochemical and mutagenesis evidence.
Supporting Evidence:
PMID:19075007
Amino acid analysis of the products of GLT25D1 and GLT25D2 reactions confirmed the transfer of galactose to hydroxylysine residues.
GO:0180062 protein O-linked glycosylation via galactose
IMP
PMID:27402836
Collagen Accumulation in Osteosarcoma Cells lacking GLT25D1 ...
ACCEPT
Summary: COLGALT1 performs the galactose-transfer step of collagen O-linked glycosylation onto hydroxylysine. Its loss reduces collagen glycosylation by up to 60% in osteosarcoma cells, directly implicating it in this process.
Reason: This accurately describes the biological process the enzyme directly carries out (O-linked galactosylation of hydroxylysine), supported by loss-of-function data.
Supporting Evidence:
PMID:27402836
Loss of GLT25D1 decreased collagen glycosylation by up to 60%.
GO:0050211 procollagen galactosyltransferase activity
IMP
PMID:27402836
Collagen Accumulation in Osteosarcoma Cells lacking GLT25D1 ...
ACCEPT
Summary: Loss of GLT25D1/COLGALT1 reduces collagen galactosylation, providing functional (mutant phenotype) support for its procollagen galactosyltransferase activity.
Reason: Core molecular function with direct loss-of-function support, corroborating the biochemical characterization.
Supporting Evidence:
PMID:27402836
Loss of GLT25D1 decreased collagen glycosylation by up to 60%.
GO:1904028 positive regulation of collagen fibril organization
IMP
PMID:27402836
Collagen Accumulation in Osteosarcoma Cells lacking GLT25D1 ...
MARK AS OVER ANNOTATED
Summary: This annotation derives from loss-of-function studies in osteosarcoma cells, but those experiments measured collagen glycosylation, intracellular collagen accumulation, and collagen type I expression, not fibril organization. The same study reported that loss of GLT25D1 did not alter collagen folding or thermal stability.
Reason: The cited paper does not demonstrate positive regulation of collagen fibril organization; it showed intracellular collagen accumulation and unchanged folding/thermal stability. Any effect on fibril organization is indirect and speculative. This regulatory term over-interprets the available evidence.
Supporting Evidence:
PMID:27402836
Loss of GLT25D1 decreased collagen glycosylation by up to 60% but did not alter collagen folding and thermal stability.
GO:0016020 membrane
HDA
PMID:19946888
Defining the membrane proteome of NK cells.
MARK AS OVER ANNOTATED
Summary: This high-throughput annotation comes from a mass-spectrometry membrane-proteome screen of NK-like YTS cells. COLGALT1 is a soluble ER-lumen protein (no transmembrane domain), and the screen itself noted that a large fraction of identified proteins were not plausible integral membrane proteins but transiently associated.
Reason: COLGALT1 is a soluble protein of the ER lumen, directly demonstrated by membrane-floatation and localization assays; it has a cleaved signal sequence and no transmembrane region. The membrane assignment reflects co-isolation in a proteomic screen rather than genuine membrane residence, and is contradicted by direct evidence.
Supporting Evidence:
PMID:20470363
The bulk of GLT25D1 is observed in the bottom fractions of the gradient, indicating the protein is not an integral membrane protein.
GO:0005788 endoplasmic reticulum lumen
TAS
Reactome:R-HSA-1981120
ACCEPT
Summary: ER lumen localization asserted by Reactome, consistent with direct experimental evidence that COLGALT1 is a soluble ER-lumen protein.
Reason: Correct site of action, corroborated by direct localization data.
Supporting Evidence:
PMID:20470363
GLT25D1 is directed to the ER lumen as a soluble protein and retained there.
GO:0005788 endoplasmic reticulum lumen
TAS
Reactome:R-HSA-8948228
ACCEPT
Summary: ER lumen localization asserted by Reactome (COLGALT1/COLGALT2 binding lysyl-hydroxylated collagen propeptides), consistent with the soluble ER-lumen localization of the enzyme.
Reason: Correct localization, corroborated by direct experimental evidence.
Supporting Evidence:
PMID:20470363
GLT25D1 is directed to the ER lumen as a soluble protein and retained there.
GO:0005788 endoplasmic reticulum lumen
TAS
Reactome:R-HSA-8948231
ACCEPT
Summary: ER lumen localization asserted by Reactome, consistent with the experimentally established soluble ER-lumen residence of COLGALT1.
Reason: Correct localization, corroborated by direct experimental evidence.
Supporting Evidence:
PMID:20470363
GLT25D1 is directed to the ER lumen as a soluble protein and retained there.
GO:0005788 endoplasmic reticulum lumen
IDA
PMID:20470363
The human collagen beta(1-O)galactosyltransferase, GLT25D1, ...
ACCEPT
Summary: Direct immunofluorescence and membrane-floatation evidence that COLGALT1 is a soluble protein of the ER lumen, retained via a C-terminal RDEL signal, co-localizing with PLOD3/LH3 and MBL in the early secretory pathway.
Reason: This is the strongest, most direct evidence for the cellular location of COLGALT1 and represents its core site of action.
Supporting Evidence:
PMID:20470363
GLT25D1 is directed to the ER lumen as a soluble protein and retained there.
GO:0030145 manganese ion binding
IC
PMID:22216269
Identification of domains and amino acids essential to the c...
NEW
Summary: Proposed annotation not present in the current GOA for COLGALT1.
Reason: COLGALT1 belongs to glycosyltransferase family 25 with a GT-A-like fold and essential catalytic aspartate residues (D166/D168, D461/D463 identified by mutagenesis), characteristic of divalent-metal (Mn2+)-dependent glycosyltransferases. Mn2+ dependence is expected on family and structural grounds, although no direct experimental metal-binding annotation currently exists in GOA.
Supporting Evidence:
PMID:22216269
Mutagenesis identified aspartate residues D166/D168 and D461/D463 as essential for collagen galactosyltransferase activity, consistent with a GT-A metal-dependent catalytic fold.

Core Functions

Collagen beta(1-O)galactosyltransferase that transfers galactose from UDP-galactose onto hydroxylysine residues of collagen in the ER lumen, catalyzing the first step of collagen O-linked glycosylation.

Supporting Evidence:
  • PMID:19075007
    Amino acid analysis of the products of GLT25D1 and GLT25D2 reactions confirmed the transfer of galactose to hydroxylysine residues.
  • PMID:27402836
    Loss of GLT25D1 decreased collagen glycosylation by up to 60%.

References

Annotation inferences using phylogenetic trees
Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping, accompanied by conservative changes to GO terms applied by UniProt
Combined Automated Annotation using Multiple IEA Methods
Core glycosylation of collagen is initiated by two beta(1-O)galactosyltransferases.
  • GLT25D1/COLGALT1 and GLT25D2 are beta(1-O)galactosyltransferases that initiate core glycosylation of collagen by transferring galactose to hydroxylysine residues; activity is the basis for EC 2.4.1.50 assignment, with KM ~18.7 uM for UDP-galactose.
Defining the membrane proteome of NK cells.
  • COLGALT1 was identified in a large-scale mass-spectrometry membrane-proteome screen of NK-like YTS cells; ~60% of identified proteins were judged not to be plausible integral membrane proteins but rather transiently membrane-associated.
The human collagen beta(1-O)galactosyltransferase, GLT25D1, is a soluble endoplasmic reticulum localized protein.
  • GLT25D1/COLGALT1 is directed to the ER lumen as a soluble protein (cleaved N-terminal signal sequence) and retained there via a C-terminal RDEL ER-retrieval signal; membrane floatation shows it is not an integral membrane protein.
  • GLT25D1 co-localizes with its substrate mannose-binding lectin (MBL) and with the upstream lysyl hydroxylase 3 (LH3/PLOD3) in the early secretory pathway.
Identification of domains and amino acids essential to the collagen galactosyltransferase activity of GLT25D1.
  • Mutagenesis identified aspartate residues D166/D168 and D461/D463 as essential for collagen galactosyltransferase activity, consistent with a GT-A metal-dependent catalytic fold; KM ~29.9 uM for UDP-galactose.
Collagen Accumulation in Osteosarcoma Cells lacking GLT25D1 Collagen Galactosyltransferase.
  • CRISPR/Cas9 inactivation of GLT25D1 in SaOS-2 osteosarcoma cells decreased collagen glycosylation by up to 60%, increased collagen type I expression, and caused intracellular (ER) accumulation of collagen type I, with compensatory induction of GLT25D2; loss did not alter collagen folding or thermal stability.
  • GLT25D1 is the main collagen galactosyltransferase isoform; GLT25D2 loss alone had no effect on collagen secretion, and clones lacking both could not be recovered, suggesting collagen glycosylation is essential for osteosarcoma cell viability.
Biallelic COLGALT1 variants are associated with cerebral small vessel disease.
  • Biallelic COLGALT1 variants (L151R, A154P, G377R) cause brain small vessel disease 3 (BSVD3); the L151R variant abolishes galactosyltransferase activity, and COLGALT1 is implicated in collagen type IV biosynthesis.
Reactome:R-HSA-1650814
Collagen biosynthesis and modifying enzymes
Reactome:R-HSA-1981120
Galactosylation of collagen propeptide hydroxylysines by procollagen galactosyltransferases 1, 2
Reactome:R-HSA-8948228
COLGALT1,COLGALT2 bind Lysyl hydroxylated collagen propeptides
Reactome:R-HSA-8948231
COLGALT1,COLGALT2:Galactosyl-hydroxylysyl collagen propeptides dissociates

Suggested Questions for Experts

Q: Does COLGALT1 require Mn2+ (or another divalent cation) for catalysis, and which residues coordinate the metal in the recently solved GT25 structures?

Q: To what extent are the collagen folding/secretion phenotypes of COLGALT1 loss tissue- and collagen-type-specific (e.g., type IV in cerebral vasculature vs. type I in bone), given paralogous compensation by COLGALT2?

Suggested Experiments

Experiment: Use the available cryo-EM and X-ray structures (e.g., PDB 8ZGE, 9EVJ) together with metal-substitution and site-directed mutagenesis to confirm the catalytic metal requirement and map the divalent-cation coordination site.

Experiment: Generate tissue-specific or vascular COLGALT1 knockouts/knock-ins of BSVD3 variants and quantify collagen type IV glycosylation, basement membrane integrity, and small-vessel pathology to causally link enzymatic loss to disease.

๐Ÿ“š Additional Documentation

Notes

(COLGALT1-notes.md)

COLGALT1 (Q8NBJ5) review notes

Identity

  • HGNC symbol: COLGALT1 (synonym GLT25D1); ORFNames PSEC0241.
  • UniProt Q8NBJ5, "Procollagen galactosyltransferase 1" / "Collagen beta(1-O)galactosyltransferase 1" (ColGalT 1) / "Glycosyltransferase 25 family member 1" / "Hydroxylysine galactosyltransferase 1".
  • EC 2.4.1.50 [ECO:0000269|PubMed:19075007].
  • CAZy GT25; glycosyltransferase 25 family.
  • 622 aa precursor; signal peptide 1-29, chain 30-622; C-terminal RDEL (619-622) ER retention motif.

Core function

  • Beta(1-O)galactosyltransferase that transfers galactose from UDP-alpha-D-galactose onto (5R)-5-hydroxy-L-lysine (hydroxylysine, Hyl) residues of collagen, forming Gal-O-Hyl. This is the FIRST step of collagen O-linked glycosylation; the resulting galactose is the acceptor for subsequent alpha1,2-glucosylation, yielding the conserved Glc(alpha1-2)Gal disaccharide [PMID:19075007; PMID:27402836 "Nascent procollagen chains are modified ... and glycosylation of selected hydroxylysine (Hyl) residues"].
  • Reaction: (5R)-5-hydroxy-L-lysyl-[collagen] + UDP-alpha-D-galactose = (5R)-5-O-(beta-D-galactosyl)-5-hydroxy-L-lysyl-[collagen] + UDP + H+ (RHEA:12637) [file:human/COLGALT1/COLGALT1-uniprot.txt CATALYTIC ACTIVITY].
  • Schegg et al. identified GLT25D1/GLT25D2 as the two beta(1-O)galactosyltransferases that initiate core glycosylation of collagen and also act on the collagenous domain of MBL [PMID:19075007 (UniProt FUNCTION/CATALYTIC ACTIVITY)].
  • KM for UDP-galactose ~18.7 uM (PubMed:19075007) / 29.9 uM (PubMed:22216269).
  • Catalytic Asp residues identified by mutagenesis: D166/D168 and D461/D463 essential for galactosyltransferase activity (loss when mutated to Ala); D-X-D-like motifs typical of metal(Mn2+)-dependent GT-A glycosyltransferases [file:human/COLGALT1/COLGALT1-uniprot.txt MUTAGEN; PubMed:22216269].

Subcellular localization

  • Soluble protein of the ENDOPLASMIC RETICULUM LUMEN. Has N-terminal cleaved signal sequence and C-terminal RDEL ER-retention/retrieval signal; membrane floatation shows it is NOT an integral membrane protein [PMID:20470363 "GLT25D1 is directed to the ER lumen as a soluble protein and retained there"; "RDEL retrieves GLT25D1 to the ER"; "The bulk of GLT25D1 is observed in the bottom fractions ... indicating the protein is not an integral membrane protein"].
  • Co-localizes with PLOD3/LH3 (lysyl hydroxylase 3, upstream enzyme) and with the substrate MBL in the early secretory pathway PMID:20470363.
  • The HDA "membrane" annotation (GO:0016020, PMID:19946888) comes from a large-scale NK-cell (YTS) membrane-proteome mass-spec study. That paper notes ~60% of identified proteins were NOT plausible integral membrane proteins but transiently membrane-associated; this is consistent with a soluble ER-lumen protein being co-isolated with membranes. Not a reliable localization claim for COLGALT1 [PMID:19946888 abstract].

Glycosyltransferase substrate / biological role

  • Loss of GLT25D1 (CRISPR/Cas9) in SaOS-2 osteosarcoma cells: decreased collagen glycosylation by up to 60%, increased collagen type I expression and intracellular (ER) accumulation, with compensatory induction of GLT25D2. GLT25D1 is the main isoform; GLT25D2 loss alone had no effect on collagen secretion. Simultaneous loss of both could not be recovered, suggesting collagen glycosylation is essential for osteosarcoma viability [PMID:27402836 abstract & Discussion]. Notably, loss of GLT25D1 "did not alter collagen folding and thermal stability" in this cell model PMID:27402836.
  • The GO IMP annotation GO:1904028 "positive regulation of collagen fibril organization" (PMID:27402836) is an over-interpretation: the paper measured intracellular collagen accumulation/expression and glycosylation, not fibril organization per se. The UniProt FUNCTION states glycosylation "facilitates the formation of collagen triple helix (PubMed:27402836)" but the paper actually reported no change in folding/thermal stability; the fibril-organization phenotype is at best indirect.

Disease

  • Biallelic COLGALT1 variants cause Brain small vessel disease 3 (BSVD3, MIM:618360), an autosomal recessive cerebral small vessel disease (porencephaly, intracranial hemorrhage, developmental delay). Variants L151R, A154P, G377R; L151R shown to cause loss of galactosyltransferase activity. Also implicated in collagen type IV biosynthesis [PMID:30412317 (UniProt FUNCTION/DISEASE)].

Structure

  • Multiple recent experimental structures (cryo-EM 8ZGC/8ZGE/8ZGG/8ZGH; X-ray 9EVJ/9EVK/9EVL), residues 30-622.
  • Tandem GT-A-like domains (HHpred hits to GT-2/pp-GalNAc-T10 and GT-31/Manic Fringe) PMID:20470363.

Annotation review summary

  • ACCEPT (core MF): GO:0050211 procollagen galactosyltransferase activity (IBA, IMP, TAS, IEA all converge; well supported experimentally).
  • ACCEPT (core CC): GO:0005788 endoplasmic reticulum lumen (IDA PMID:20470363; multiple TAS/IEA).
  • ACCEPT / KEEP (BP): GO:0180062 protein O-linked glycosylation via galactose (IMP) โ€” accurate description of the process the enzyme performs.
  • KEEP_AS_NON_CORE: GO:0030199 collagen fibril organization (TAS Reactome) โ€” downstream/indirect.
  • MARK_AS_OVER_ANNOTATED: GO:1904028 positive regulation of collagen fibril organization (IMP PMID:27402836) โ€” over-interpretation of the loss-of-function data.
  • MARK_AS_OVER_ANNOTATED: GO:0016020 membrane (HDA PMID:19946888) โ€” soluble ER-lumen protein co-isolated in a membrane proteome screen; contradicted by IDA soluble-ER evidence.

Proposed additions (not in current GOA)

  • GO:0030145 manganese ion binding โ€” UniProt/EC and GT-A-fold D-X-D catalytic Asp residues (D461/D463 essential by mutagenesis) indicate Mn2+-dependence typical of GT-A glycosyltransferases; no direct experimental metal-binding annotation exists in GOA. Proposed as a new term on family/structural grounds.

Pn Notes

(COLGALT1-pn-notes.md)

COLGALT1 PN Consistency Notes

  • Generated: 2026-06-18
  • Project: PROTEOSTASIS
  • Scope: PN consistency rereview against local AIGR review and available deep-research artifacts
  • UniProt: Q8NBJ5
  • AIGR review status: COMPLETE
  • Review batch: proteostasis-batch-2026-06-07
  • Batch change status: added

Source Files Checked

Deep Research Files

  • No *-deep-research*.md file found in this gene directory.

AIGR Review Snapshot

  • Description: COLGALT1 (collagen beta(1-O)galactosyltransferase 1; also known as GLT25D1) is a soluble enzyme of the endoplasmic reticulum lumen that catalyzes the first committed step of collagen O-linked glycosylation. It transfers galactose from UDP-alpha-D-galactose onto (5R)-5-hydroxy-L-lysine (hydroxylysine) residues of collagen, forming the beta(1-O)-linked Gal-O-hydroxylysine (EC 2.4.1.50). This galactose is then the acceptor for subsequent alpha1,2-glucosylation, generating the Glc(alpha1-2)Gal disaccharide that is strongly conserved across animal collagens. The enzyme belongs to glycosyltransferase family 25 (GT25), adopts a GT-A-like fold with metal-dependent (Mn2+) catalysis, and uses essential aspartate residues (D166/D168, D461/D463) for activity. COLGALT1 acts on multiple collagen types (including types I and IV) as well as the collagenous domain of mannose-binding lectin, and co-localizes in the early secretory pathway with the upstream lysyl hydroxylase PLOD3/LH3. As the predominant of two paralogous collagen galactosyltransferases (the other being COLGALT2/GLT25D2), it is broadly expressed and its loss reduces collagen glycosylation, causing intracellular collagen accumulation. Biallelic loss-of-function variants in COLGALT1 cause an autosomal recessive cerebral small vessel disease (brain small vessel disease 3).
  • Existing/core annotation action counts: ACCEPT: 10; KEEP_AS_NON_CORE: 1; MARK_AS_OVER_ANNOTATED: 2; NEW: 1

PN Consistency Summary

  • Consistency: Consistent. Review, notes-equivalent (rich references), PN annotation, and PN-node mapping all describe COLGALT1/GLT25D1 as a soluble ER-lumen collagen beta(1-O)galactosyltransferase (EC 2.4.1.50) performing the first committed step of collagen O-glycosylation. The collagen-biosynthesis framing is concordant across sources.
  • PN story / NEW pressure: PN projects collagen biosynthetic process (GO:0032964, verified real OLS). COLGALT1's curated core function is the more specific GO:0050211 procollagen galactosyltransferase activity (MF) plus GO:0180062 protein O-linked glycosylation via galactose (BP), and it already carries GO:0030199 collagen fibril organization. GO:0032964 is a broader pathway-context process consistent with these but more general. Conclusion: PN context is already captured at a finer grain in the review; no additional NEW-term pressure from the PN node. (Review's own NEW is GO:0030145 Mn ion binding โ€” orthogonal to PN.)
  • Evidence alignment: PN row carries no reference titles; review evidence (PMID:19075007, PMID:20470363, PMID:22216269, PMID:27402836, PMID:30412317, Reactome collagen-galactosylation reactions) fully supports the collagen-biosynthesis mapping. No divergence.
  • Verdict: Consistent; PN collagen-biosynthesis role already captured (more specifically) in review. No edits required.

Full Consistency Review

  • UniProt: Q8NBJ5 ยท batch: proteostasis-batch-2026-06-07 ยท review status: COMPLETE
  • PN placement: ER proteostasis | Maturation and folding of specific substrates | ER collagen processing and folding ; PN-node mapping: group=mapped, scope=ok_for_propagation_to_go, GO:0032964 collagen biosynthetic process (class/branch = no_mapping)
  • Consistency: Consistent. Review, notes-equivalent (rich references), PN annotation, and PN-node mapping all describe COLGALT1/GLT25D1 as a soluble ER-lumen collagen beta(1-O)galactosyltransferase (EC 2.4.1.50) performing the first committed step of collagen O-glycosylation. The collagen-biosynthesis framing is concordant across sources.
  • PN story / NEW pressure: PN projects collagen biosynthetic process (GO:0032964, verified real OLS). COLGALT1's curated core function is the more specific GO:0050211 procollagen galactosyltransferase activity (MF) plus GO:0180062 protein O-linked glycosylation via galactose (BP), and it already carries GO:0030199 collagen fibril organization. GO:0032964 is a broader pathway-context process consistent with these but more general. Conclusion: PN context is already captured at a finer grain in the review; no additional NEW-term pressure from the PN node. (Review's own NEW is GO:0030145 Mn ion binding โ€” orthogonal to PN.)
  • Mapping strategy: No change. Mapping the group node to GO:0032964 is appropriate as a shared substrate-specific pathway context for the collagen-processing group; it is broader than COLGALT1's specific MF/BP but is a process-level umbrella, not an over-reach (unlike a CC/complex claim). Ancestors correctly no_mapping.
  • Evidence alignment: PN row carries no reference titles; review evidence (PMID:19075007, PMID:20470363, PMID:22216269, PMID:27402836, PMID:30412317, Reactome collagen-galactosylation reactions) fully supports the collagen-biosynthesis mapping. No divergence.
  • Verdict: Consistent; PN collagen-biosynthesis role already captured (more specifically) in review. No edits required.

PN Dossier Context

  • review_batch: proteostasis-batch-2026-06-07
  • review_yaml: genes/human/COLGALT1/COLGALT1-ai-review.yaml
  • PN workbook rows: 1

PN row 1: ER proteostasis | Maturation and folding of specific substrates | ER collagen processing and folding

  • UniProt: Q8NBJ5
  • In branches: ER
  • PN-node mapping records (path + ancestors):
    • [group] ER proteostasis|Maturation and folding of specific substrates|ER collagen processing and folding
      status=mapped scope=ok_for_propagation_to_go GO=[GO:0032964 collagen biosynthetic process]
      rationale: This PN group contains ER factors dedicated to collagen maturation, processing, and folding. Collagen biosynthetic process captures the shared substrate-specific pathway context.
    • [class] ER proteostasis|Maturation and folding of specific substrates
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a broad PN category rather than a single GO class. The member genes span multiple activities, complexes, or contexts, so direct propagation from this node would overstate the shared biology.
    • [branch] ER proteostasis
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a top-level PN branch. This is a systems/taxonomy umbrella, not a direct GO assertion; narrower child curations carry any propagating GO mappings.

Projected GO annotations (1)

  • GO:0032964 collagen biosynthetic process | scope=ok_for_propagation_to_go | goa_status=new_to_goa | from=ER proteostasis|Maturation and folding of specific substrates|ER collagen processing and folding

Note

This file is generated from the current PROTEOSTASIS phase-1 dossier and local gene-review artifacts. Edit the source review, PN mapping, or dossier rather than this generated note when correcting the underlying curation.

๐Ÿ“„ View Raw YAML

id: Q8NBJ5
gene_symbol: COLGALT1
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: COLGALT1 (collagen beta(1-O)galactosyltransferase 1; also known as GLT25D1)
  is a soluble enzyme of the endoplasmic reticulum lumen that catalyzes the first
  committed step of collagen O-linked glycosylation. It transfers galactose from UDP-alpha-D-galactose
  onto (5R)-5-hydroxy-L-lysine (hydroxylysine) residues of collagen, forming the beta(1-O)-linked
  Gal-O-hydroxylysine (EC 2.4.1.50). This galactose is then the acceptor for subsequent
  alpha1,2-glucosylation, generating the Glc(alpha1-2)Gal disaccharide that is strongly
  conserved across animal collagens. The enzyme belongs to glycosyltransferase family
  25 (GT25), adopts a GT-A-like fold with metal-dependent (Mn2+) catalysis, and uses
  essential aspartate residues (D166/D168, D461/D463) for activity. COLGALT1 acts on
  multiple collagen types (including types I and IV) as well as the collagenous domain
  of mannose-binding lectin, and co-localizes in the early secretory pathway with the
  upstream lysyl hydroxylase PLOD3/LH3. As the predominant of two paralogous collagen
  galactosyltransferases (the other being COLGALT2/GLT25D2), it is broadly expressed
  and its loss reduces collagen glycosylation, causing intracellular collagen accumulation.
  Biallelic loss-of-function variants in COLGALT1 cause an autosomal recessive cerebral
  small vessel disease (brain small vessel disease 3).
references:
- id: GO_REF:0000033
  title: Annotation inferences using phylogenetic trees
  findings: []
- id: GO_REF:0000044
  title: Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location
    vocabulary mapping, accompanied by conservative changes to GO terms applied by
    UniProt
  findings: []
- id: GO_REF:0000120
  title: Combined Automated Annotation using Multiple IEA Methods
  findings: []
- id: PMID:19075007
  title: Core glycosylation of collagen is initiated by two beta(1-O)galactosyltransferases.
  findings:
  - statement: GLT25D1/COLGALT1 and GLT25D2 are beta(1-O)galactosyltransferases that
      initiate core glycosylation of collagen by transferring galactose to hydroxylysine
      residues; activity is the basis for EC 2.4.1.50 assignment, with KM ~18.7 uM
      for UDP-galactose.
    reference_section_type: RESULTS
- id: PMID:19946888
  title: Defining the membrane proteome of NK cells.
  findings:
  - statement: COLGALT1 was identified in a large-scale mass-spectrometry membrane-proteome
      screen of NK-like YTS cells; ~60% of identified proteins were judged not to be
      plausible integral membrane proteins but rather transiently membrane-associated.
    reference_section_type: ABSTRACT
- id: PMID:20470363
  title: The human collagen beta(1-O)galactosyltransferase, GLT25D1, is a soluble
    endoplasmic reticulum localized protein.
  findings:
  - statement: GLT25D1/COLGALT1 is directed to the ER lumen as a soluble protein (cleaved
      N-terminal signal sequence) and retained there via a C-terminal RDEL ER-retrieval
      signal; membrane floatation shows it is not an integral membrane protein.
    reference_section_type: RESULTS
  - statement: GLT25D1 co-localizes with its substrate mannose-binding lectin (MBL)
      and with the upstream lysyl hydroxylase 3 (LH3/PLOD3) in the early secretory
      pathway.
    reference_section_type: RESULTS
- id: PMID:22216269
  title: Identification of domains and amino acids essential to the collagen galactosyltransferase
    activity of GLT25D1.
  findings:
  - statement: Mutagenesis identified aspartate residues D166/D168 and D461/D463 as
      essential for collagen galactosyltransferase activity, consistent with a GT-A
      metal-dependent catalytic fold; KM ~29.9 uM for UDP-galactose.
    reference_section_type: RESULTS
- id: PMID:27402836
  title: Collagen Accumulation in Osteosarcoma Cells lacking GLT25D1 Collagen Galactosyltransferase.
  findings:
  - statement: CRISPR/Cas9 inactivation of GLT25D1 in SaOS-2 osteosarcoma cells decreased
      collagen glycosylation by up to 60%, increased collagen type I expression, and
      caused intracellular (ER) accumulation of collagen type I, with compensatory
      induction of GLT25D2; loss did not alter collagen folding or thermal stability.
    reference_section_type: RESULTS
  - statement: GLT25D1 is the main collagen galactosyltransferase isoform; GLT25D2
      loss alone had no effect on collagen secretion, and clones lacking both could
      not be recovered, suggesting collagen glycosylation is essential for osteosarcoma
      cell viability.
    reference_section_type: DISCUSSION
- id: PMID:30412317
  title: Biallelic COLGALT1 variants are associated with cerebral small vessel disease.
  findings:
  - statement: Biallelic COLGALT1 variants (L151R, A154P, G377R) cause brain small
      vessel disease 3 (BSVD3); the L151R variant abolishes galactosyltransferase
      activity, and COLGALT1 is implicated in collagen type IV biosynthesis.
    reference_section_type: RESULTS
- id: Reactome:R-HSA-1650814
  title: Collagen biosynthesis and modifying enzymes
  findings: []
- id: Reactome:R-HSA-1981120
  title: Galactosylation of collagen propeptide hydroxylysines by procollagen galactosyltransferases
    1, 2
  findings: []
- id: Reactome:R-HSA-8948228
  title: COLGALT1,COLGALT2 bind Lysyl hydroxylated collagen propeptides
  findings: []
- id: Reactome:R-HSA-8948231
  title: COLGALT1,COLGALT2:Galactosyl-hydroxylysyl collagen propeptides dissociates
  findings: []
existing_annotations:
- term:
    id: GO:0050211
    label: procollagen galactosyltransferase activity
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: Phylogenetically inferred procollagen galactosyltransferase activity.
      This is the well-established core molecular function of COLGALT1, directly demonstrated
      experimentally for the human enzyme.
    action: ACCEPT
    reason: This is the core molecular function of COLGALT1, supported by direct biochemical
      characterization (EC 2.4.1.50) and mutagenesis. The IBA transfer is fully consistent
      with the experimental evidence.
    supported_by:
    - reference_id: PMID:19075007
      supporting_text: >-
        Amino acid analysis of the products of GLT25D1 and GLT25D2 reactions confirmed the
        transfer of galactose to hydroxylysine residues.
- term:
    id: GO:0005788
    label: endoplasmic reticulum lumen
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: ER lumen localization mapped from UniProt subcellular location. COLGALT1
      is a soluble ER-lumen protein, directly shown by immunofluorescence and membrane-floatation
      assays.
    action: ACCEPT
    reason: ER lumen is the correct site of action; this electronic mapping agrees
      with direct experimental localization evidence.
    supported_by:
    - reference_id: PMID:20470363
      supporting_text: >-
        In agreement with the predictions our results show that GLT25D1 is directed to the ER
        lumen as a soluble protein and retained there.
- term:
    id: GO:0050211
    label: procollagen galactosyltransferase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: Electronically inferred procollagen galactosyltransferase activity (mapped
      from RHEA:12637 / EC 2.4.1.50). This is the experimentally validated core function.
    action: ACCEPT
    reason: Consistent with the experimentally demonstrated catalytic activity and
      EC assignment.
    supported_by:
    - reference_id: file:human/COLGALT1/COLGALT1-uniprot.txt
      supporting_text: EC=2.4.1.50; Reaction=(5R)-5-hydroxy-L-lysyl-[collagen] + UDP-alpha-D-galactose
        = (5R)-5-O-(beta-D-galactosyl)-5-hydroxy-L-lysyl-[collagen] + UDP + H(+); Xref=Rhea:RHEA:12637.
- term:
    id: GO:0030199
    label: collagen fibril organization
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-1650814
  qualifier: involved_in
  review:
    summary: COLGALT1 contributes to collagen biosynthesis/modification, which is upstream
      of collagen fibril organization in the extracellular matrix. Its direct action
      is intracellular hydroxylysine galactosylation, not fibril assembly itself.
    action: KEEP_AS_NON_CORE
    reason: This is a downstream, indirect biological consequence of collagen glycosylation
      rather than the core molecular function. Glycosylation has been suggested to
      influence collagen crosslink formation and matrix organization, but the enzyme
      does not directly organize fibrils. Retaining as non-core captures the pathway
      context without overstating the role.
    supported_by:
    - reference_id: PMID:27402836
      supporting_text: glycosylation may be involved in the organization of collagens
        in the extracellular space. Glycosylation has been suggested to regulate cross-link
        formation in fibrillar collagens.
- term:
    id: GO:0050211
    label: procollagen galactosyltransferase activity
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-1981120
  qualifier: enables
  review:
    summary: Reactome-asserted procollagen galactosyltransferase activity, the core
      molecular function of COLGALT1.
    action: ACCEPT
    reason: Core molecular function, well supported by direct biochemical and mutagenesis
      evidence.
    supported_by:
    - reference_id: PMID:19075007
      supporting_text: >-
        Amino acid analysis of the products of GLT25D1 and GLT25D2 reactions confirmed the
        transfer of galactose to hydroxylysine residues.
- term:
    id: GO:0180062
    label: protein O-linked glycosylation via galactose
  evidence_type: IMP
  original_reference_id: PMID:27402836
  qualifier: involved_in
  review:
    summary: COLGALT1 performs the galactose-transfer step of collagen O-linked glycosylation
      onto hydroxylysine. Its loss reduces collagen glycosylation by up to 60% in osteosarcoma
      cells, directly implicating it in this process.
    action: ACCEPT
    reason: This accurately describes the biological process the enzyme directly carries
      out (O-linked galactosylation of hydroxylysine), supported by loss-of-function
      data.
    supported_by:
    - reference_id: PMID:27402836
      supporting_text: Loss of GLT25D1 decreased collagen glycosylation by up to 60%.
- term:
    id: GO:0050211
    label: procollagen galactosyltransferase activity
  evidence_type: IMP
  original_reference_id: PMID:27402836
  qualifier: enables
  review:
    summary: Loss of GLT25D1/COLGALT1 reduces collagen galactosylation, providing functional
      (mutant phenotype) support for its procollagen galactosyltransferase activity.
    action: ACCEPT
    reason: Core molecular function with direct loss-of-function support, corroborating
      the biochemical characterization.
    supported_by:
    - reference_id: PMID:27402836
      supporting_text: Loss of GLT25D1 decreased collagen glycosylation by up to 60%.
- term:
    id: GO:1904028
    label: positive regulation of collagen fibril organization
  evidence_type: IMP
  original_reference_id: PMID:27402836
  qualifier: involved_in
  review:
    summary: This annotation derives from loss-of-function studies in osteosarcoma cells,
      but those experiments measured collagen glycosylation, intracellular collagen
      accumulation, and collagen type I expression, not fibril organization. The same
      study reported that loss of GLT25D1 did not alter collagen folding or thermal
      stability.
    action: MARK_AS_OVER_ANNOTATED
    reason: The cited paper does not demonstrate positive regulation of collagen fibril
      organization; it showed intracellular collagen accumulation and unchanged folding/thermal
      stability. Any effect on fibril organization is indirect and speculative. This
      regulatory term over-interprets the available evidence.
    supported_by:
    - reference_id: PMID:27402836
      supporting_text: Loss of GLT25D1 decreased collagen glycosylation by up to 60%
        but did not alter collagen folding and thermal stability.
- term:
    id: GO:0016020
    label: membrane
  evidence_type: HDA
  original_reference_id: PMID:19946888
  qualifier: located_in
  review:
    summary: This high-throughput annotation comes from a mass-spectrometry membrane-proteome
      screen of NK-like YTS cells. COLGALT1 is a soluble ER-lumen protein (no transmembrane
      domain), and the screen itself noted that a large fraction of identified proteins
      were not plausible integral membrane proteins but transiently associated.
    action: MARK_AS_OVER_ANNOTATED
    reason: COLGALT1 is a soluble protein of the ER lumen, directly demonstrated by
      membrane-floatation and localization assays; it has a cleaved signal sequence
      and no transmembrane region. The membrane assignment reflects co-isolation in
      a proteomic screen rather than genuine membrane residence, and is contradicted
      by direct evidence.
    supported_by:
    - reference_id: PMID:20470363
      supporting_text: The bulk of GLT25D1 is observed in the bottom fractions of the
        gradient, indicating the protein is not an integral membrane protein.
- term:
    id: GO:0005788
    label: endoplasmic reticulum lumen
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-1981120
  qualifier: located_in
  review:
    summary: ER lumen localization asserted by Reactome, consistent with direct experimental
      evidence that COLGALT1 is a soluble ER-lumen protein.
    action: ACCEPT
    reason: Correct site of action, corroborated by direct localization data.
    supported_by:
    - reference_id: PMID:20470363
      supporting_text: GLT25D1 is directed to the ER lumen as a soluble protein and
        retained there.
- term:
    id: GO:0005788
    label: endoplasmic reticulum lumen
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8948228
  qualifier: located_in
  review:
    summary: ER lumen localization asserted by Reactome (COLGALT1/COLGALT2 binding lysyl-hydroxylated
      collagen propeptides), consistent with the soluble ER-lumen localization of the
      enzyme.
    action: ACCEPT
    reason: Correct localization, corroborated by direct experimental evidence.
    supported_by:
    - reference_id: PMID:20470363
      supporting_text: GLT25D1 is directed to the ER lumen as a soluble protein and
        retained there.
- term:
    id: GO:0005788
    label: endoplasmic reticulum lumen
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8948231
  qualifier: located_in
  review:
    summary: ER lumen localization asserted by Reactome, consistent with the experimentally
      established soluble ER-lumen residence of COLGALT1.
    action: ACCEPT
    reason: Correct localization, corroborated by direct experimental evidence.
    supported_by:
    - reference_id: PMID:20470363
      supporting_text: GLT25D1 is directed to the ER lumen as a soluble protein and
        retained there.
- term:
    id: GO:0005788
    label: endoplasmic reticulum lumen
  evidence_type: IDA
  original_reference_id: PMID:20470363
  qualifier: located_in
  review:
    summary: Direct immunofluorescence and membrane-floatation evidence that COLGALT1
      is a soluble protein of the ER lumen, retained via a C-terminal RDEL signal,
      co-localizing with PLOD3/LH3 and MBL in the early secretory pathway.
    action: ACCEPT
    reason: This is the strongest, most direct evidence for the cellular location of
      COLGALT1 and represents its core site of action.
    supported_by:
    - reference_id: PMID:20470363
      supporting_text: GLT25D1 is directed to the ER lumen as a soluble protein and
        retained there.
- term:
    id: GO:0030145
    label: manganese ion binding
  evidence_type: IC
  original_reference_id: PMID:22216269
  qualifier: enables
  review:
    summary: Proposed annotation not present in the current GOA for COLGALT1.
    action: NEW
    reason: COLGALT1 belongs to glycosyltransferase family 25 with a GT-A-like fold and 
      essential catalytic aspartate residues (D166/D168, D461/D463 identified by 
      mutagenesis), characteristic of divalent-metal (Mn2+)-dependent 
      glycosyltransferases. Mn2+ dependence is expected on family and structural 
      grounds, although no direct experimental metal-binding annotation currently exists
      in GOA.
    supported_by:
    - reference_id: PMID:22216269
      supporting_text: Mutagenesis identified aspartate residues D166/D168 and D461/D463
        as essential for collagen galactosyltransferase activity, consistent with a GT-A
        metal-dependent catalytic fold.
      full_text_unavailable: true
core_functions:
- description: Collagen beta(1-O)galactosyltransferase that transfers galactose from
    UDP-galactose onto hydroxylysine residues of collagen in the ER lumen, catalyzing
    the first step of collagen O-linked glycosylation.
  supported_by:
  - reference_id: PMID:19075007
    supporting_text: >-
      Amino acid analysis of the products of GLT25D1 and GLT25D2 reactions confirmed the
      transfer of galactose to hydroxylysine residues.
  - reference_id: PMID:27402836
    supporting_text: Loss of GLT25D1 decreased collagen glycosylation by up to 60%.
  molecular_function:
    id: GO:0050211
    label: procollagen galactosyltransferase activity
  directly_involved_in:
  - id: GO:0180062
    label: protein O-linked glycosylation via galactose
  locations:
  - id: GO:0005788
    label: endoplasmic reticulum lumen
proposed_new_terms: []
suggested_questions:
- question: Does COLGALT1 require Mn2+ (or another divalent cation) for catalysis,
    and which residues coordinate the metal in the recently solved GT25 structures?
- question: To what extent are the collagen folding/secretion phenotypes of COLGALT1
    loss tissue- and collagen-type-specific (e.g., type IV in cerebral vasculature
    vs. type I in bone), given paralogous compensation by COLGALT2?
suggested_experiments:
- description: Use the available cryo-EM and X-ray structures (e.g., PDB 8ZGE, 9EVJ) together with metal-substitution and site-directed mutagenesis to confirm the catalytic metal requirement and map the divalent-cation coordination site.
- description: Generate tissue-specific or vascular COLGALT1 knockouts/knock-ins of BSVD3 variants and quantify collagen type IV glycosylation, basement membrane integrity, and small-vessel pathology to causally link enzymatic loss to disease.