GO:0140563 UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0140563 describes the enzymatic activity that transfers xylose from UDP-alpha-D-xylose onto a glucose residue attached to serine within EGF-like domains, forming an alpha-1,3 linkage.
• This activity is required for biosynthesis of the Xyl alpha1-3Glc beta-Ser and Xyl alpha1-3Xyl alpha1-3Glc beta1-O-Ser structures on glycoproteins.
• The enzyme was first detected in the human hepatoma cell line HepG2 and later purified and characterized for substrate specificity.
• It acts on EGF-like domain substrates and is distinct from other xylosyltransferases that modify different acceptor sites.
• Loss or alteration of this activity can change the structure of EGF-like domain glycans, which may affect protein function and interactions.
• Researchers study GO:0140563 using enzymatic assays, glycoproteomics, and CRISPR-based models to dissect its role in glycoprotein biology.
Description
GO:0140563, UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity, is a molecular function that catalyzes the transfer of xylose from UDP-alpha-D-xylose to a beta-glucosyl residue on a serine within EGF-like domains, creating an alpha-1,3 linkage. This modification is part of the biosynthesis of specific O-linked glycan structures found on glycoproteins, such as Xyl alpha1-3Glc beta-Ser and Xyl alpha1-3Xyl alpha1-3Glc beta1-O-Ser. The activity was first identified in the human hepatoma cell line HepG2, providing evidence for its presence in human cells. Subsequent purification and substrate specificity studies confirmed its preference for EGF-like domain acceptors. Understanding this enzyme is important because EGF-like domain glycosylation can influence protein folding, stability, and interactions, with implications for cell signaling and disease.
UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity At A Glance
| GO ID | GO:0140563 |
|---|---|
| GO term | UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity |
| Ontology | molecular_function |
| Synonym | None |
| Major function | Catalyzes xylose transfer to beta-glucosyl-serine in EGF-like domains |
| Substrate donor | UDP-alpha-D-xylose |
| Acceptor | Protein with EGF-like domain-3-O-(beta-D-glucosyl)-L-serine |
| Product | Protein with EGF-like domain-3-O-(alpha-D-xylosyl-(1->3)-beta-D-glucosyl)-L-serine and UDP |
| Cellular context | Glycoprotein biosynthesis in the secretory pathway |
What Is GO:0140563?
This term describes an enzyme activity that uses UDP-alpha-D-xylose as a donor to add a xylose molecule to a beta-D-glucosyl group attached to a serine residue within a protein that contains EGF-like domains. The reaction forms an alpha-1,3 linkage between xylose and glucose, producing a specific O-linked glycan structure and releasing UDP.
Why Is UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity Important in Cell Biology?
GO:0140563 is important because it defines a specific step in the biosynthesis of O-linked glycans on EGF-like domains, which are found in many cell surface and secreted proteins. This modification can affect protein function, stability, and interactions, and its dysregulation has been linked to altered glycoprotein biology in cancer and other diseases.
• Defines a key enzymatic step in O-linked glycosylation of EGF-like domains.
• Required for the formation of Xyl alpha1-3Glc beta-Ser and Xyl alpha1-3Xyl alpha1-3Glc beta1-O-Ser structures.
• Provides a molecular marker for studying glycoprotein biosynthesis in human cells.
• Enables research on the role of EGF-like domain glycans in cell signaling.
• Potential target for modulating glycoprotein function in disease.
• Supports development of enzymatic assays for drug discovery.
• Helps understand substrate specificity within the xylosyltransferase family.
• Facilitates glycoproteomic studies of EGF-like domain modifications.
What Happens During UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity?
Substrate recognition and binding
In simple terms: The enzyme finds and grabs its starting materials.
The enzyme binds UDP-alpha-D-xylose as the donor substrate and a protein containing an EGF-like domain with a beta-glucosyl-serine as the acceptor. Substrate specificity studies indicate that the enzyme recognizes the EGF-like domain structure and the glucose moiety for efficient catalysis.
Catalytic transfer of xylose
In simple terms: The enzyme moves xylose from the donor to the acceptor.
The enzyme catalyzes the transfer of xylose from UDP-alpha-D-xylose to the beta-glucosyl residue, forming an alpha-1,3 linkage and releasing UDP. This reaction is part of the biosynthesis of the Xyl alpha1-3Glc beta-Ser structure.
Product formation and elongation
In simple terms: The new sugar structure can be further modified.
The product, a protein with EGF-like domain-3-O-(alpha-D-xylosyl-(1->3)-beta-D-glucosyl)-L-serine, can serve as a substrate for further xylosylation to form Xyl alpha1-3Xyl alpha1-3Glc beta1-O-Ser.
Key Genes Involved in GO:0140563 UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity
The following genes and proteins are directly or indirectly associated with GO:0140563 activity.
| Gene | Major Role | Research Relevance |
|---|---|---|
| XYLT1 | Xylosyltransferase that may initiate glycosaminoglycan biosynthesis | Potential related enzyme in xylose transfer pathways |
| XYLT2 | Xylosyltransferase involved in proteoglycan synthesis | May share substrate overlap with GO:0140563 |
| B4GALT | Beta-1,4-galactosyltransferase family | May create beta-glucosyl acceptors for GO:0140563 |
| POGLUT1 | Protein O-glucosyltransferase that adds glucose to EGF-like domains | Generates the beta-glucosyl-serine acceptor for GO:0140563 |
| XXYLT1 | Xyloside xylosyltransferase that adds xylose to glucose | May cooperate with GO:0140563 in glycan elongation |
| EGF | Epidermal growth factor containing EGF-like domains | Model substrate for studying GO:0140563 |
| NOTCH1 | Notch receptor with EGF-like domains | Potential target of GO:0140563 glycosylation |
| NOTCH2 | Notch receptor with EGF-like domains | Potential target of GO:0140563 glycosylation |
| NOTCH3 | Notch receptor with EGF-like domains | Potential target of GO:0140563 glycosylation |
| NOTCH4 | Notch receptor with EGF-like domains | Potential target of GO:0140563 glycosylation |
| FBN1 | Fibrillin-1 with EGF-like domains | Potential substrate for GO:0140563 |
| FBN2 | Fibrillin-2 with EGF-like domains | Potential substrate for GO:0140563 |
| LTBP1 | Latent TGF-beta binding protein with EGF-like domains | Potential substrate for GO:0140563 |
| LTBP2 | Latent TGF-beta binding protein with EGF-like domains | Potential substrate for GO:0140563 |
| JAG1 | Jagged-1 with EGF-like domains | Potential substrate for GO:0140563 |
| JAG2 | Jagged-2 with EGF-like domains | Potential substrate for GO:0140563 |
| DLL1 | Delta-like ligand with EGF-like domains | Potential substrate for GO:0140563 |
How Is UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity Regulated?
The activity of GO:0140563 may be regulated by the availability of UDP-alpha-D-xylose and the presence of appropriate EGF-like domain substrates. Substrate specificity studies suggest that the enzyme recognizes specific structural features of the acceptor, which could be a point of regulation.
UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| NOTCH1 | Notch signaling in cancer | Knockout of GO:0140563 enzyme in cancer cell lines |
| FBN1 | Marfan syndrome | Point mutation in EGF-like domain to alter glycosylation |
| XYLT1 | Desbuquois dysplasia | Knock-in of patient mutations |
| POGLUT1 | Muscular dystrophy | Overexpression of GO:0140563 enzyme |
| JAG1 | Alagille syndrome | Knockout of GO:0140563 enzyme in model organisms |
Cancer
Altered glycosylation of EGF-like domains, including modifications by GO:0140563, may contribute to cancer progression by affecting cell signaling pathways. The enzyme was initially identified in a hepatoma cell line, suggesting a potential role in liver cancer biology.
Notch signaling disorders
Notch receptors contain EGF-like domains that can be modified by xylosylation. Changes in GO:0140563 activity could impact Notch signaling, which is involved in developmental disorders and cancer.
Connective tissue disorders
Proteins such as fibrillins and latent TGF-beta binding proteins contain EGF-like domains. Glycosylation by GO:0140563 may influence their function, with potential implications for connective tissue diseases.
From UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| What is the role of GO:0140563 in Notch signaling? | Knockout of the enzyme in cell lines |
| How does xylosylation affect EGF-like domain function? | Point mutation of acceptor serine |
| Can xylosylation be tracked in live cells? | Knock-in of tagged enzyme |
| Does overexpression alter glycan structures? | Overexpression of the enzyme |
| Which proteins are substrates? | Knockout followed by glycoproteomics |
| Is the enzyme required for development? | Knockout in model organisms |
How to Study the UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Enzymatic assay | Xylosyltransferase activity | Kinetic studies |
| Mass spectrometry | Glycan structures | Substrate identification |
| CRISPR knockout | Gene function | Loss-of-function studies |
| Overexpression | Gain-of-function | Phenotype analysis |
| RNA-seq | Transcriptional changes | Pathway analysis |
| Proteomics | Protein interactions | Complex identification |
| Imaging | Subcellular localization | Trafficking studies |
Enzymatic assays
In vitro assays using UDP-alpha-D-xylose and EGF-like domain substrates can measure GO:0140563 activity directly.
Glycoproteomics
Mass spectrometry-based glycoproteomics can identify xylosylated EGF-like domains and quantify changes upon enzyme manipulation.
CRISPR screening
Genome-wide CRISPR screens can identify genes that regulate GO:0140563 activity or its substrates.
Cell-based imaging
Fluorescently tagged enzymes or substrates can visualize the subcellular localization of GO:0140563 activity.
How CRISPR Can Be Used to Study GO:0140563 UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity
Knockout
CRISPR knockout of the gene encoding GO:0140563 can abolish xylosylation of EGF-like domains, allowing researchers to study its loss-of-function phenotypes.
Point Mutation
Introducing point mutations in the catalytic domain or substrate-binding residues can dissect the enzymatic mechanism of GO:0140563.
Knock-in
Knock-in of tagged or reporter versions of the enzyme enables tracking of its expression and localization in cells.
Overexpression
Overexpression of the enzyme can increase xylosylation of EGF-like domains, helping to identify downstream effects.
How EDITGENE Supports UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity Research
Researchers studying UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in glycosylation pathways. EDITGENE provides CRISPR-based tools to create precise cellular models for such investigations.
Contact EDITGENE today to design your custom CRISPR model for UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity research.
Frequently Asked Questions About UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity
What is GO:0140563?
GO:0140563 is a molecular function term for the enzyme activity that transfers xylose from UDP-alpha-D-xylose to a beta-glucosyl residue on EGF-like domains.
What genes are involved in UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase activity?
Genes such as POGLUT1, XXYLT1, and NOTCH family members are related to this activity.
What is the function of UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase?
It catalyzes the formation of an alpha-1,3 linkage between xylose and glucose on EGF-like domains, contributing to O-linked glycan biosynthesis.
Which diseases are associated with this activity?
Altered activity may impact cancer, Notch signaling disorders, and connective tissue diseases.
How can I study GO:0140563 in the lab?
Enzymatic assays, glycoproteomics, and CRISPR knockout models are common approaches.
What substrates does this enzyme use?
It uses UDP-alpha-D-xylose as donor and a protein with EGF-like domain-3-O-(beta-D-glucosyl)-L-serine as acceptor.
Is this enzyme found in humans?
Yes, it was first identified in human HepG2 cells.
What is the product of this enzymatic reaction?
The product is a protein with EGF-like domain-3-O-(alpha-D-xylosyl-(1->3)-beta-D-glucosyl)-L-serine and UDP.
Can CRISPR be used to study this activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are valuable for studying GO:0140563.
What methods measure this activity?
Enzymatic assays and mass spectrometry are commonly used to measure GO:0140563 activity.
Conclusion
GO:0140563 defines a specific xylosyltransferase activity critical for modifying EGF-like domains on glycoproteins. Its study provides insights into glycoprotein biosynthesis and related diseases, and CRISPR-based models offer powerful tools for functional dissection.
References
- 1. Omichi K et al.. 1997. Presence of UDP-D-xylose: beta-D-glucoside alpha-1,3-D-xylosyltransferase involved in the biosynthesis of the Xyl alpha 1-3Glc beta-Ser structure of glycoproteins in the human hepatoma cell line HepG2.. Eur J Biochem 245(1):143-6 PMID: 9128735
- 2. Ishimizu T et al.. 2007. Purification and substrate specificity of UDP-D-xylose:beta-D-glucoside alpha-1,3-D-xylosyltransferase involved in the biosynthesis of the Xyl alpha1-3Xyl alpha1-3Glc beta1-O-Ser on epidermal growth factor-like domains.. J Biochem 141(4):593-600 PMID: 17317689
