Disulfide Bond Formation and N-Glycosylation Modulate Protein-Protein Interactions in GPI-Transamidase (GPIT)
Open accessAbstract
Glycosylphosphatidylinositol (GPI) transamidase (GPIT), the enzyme that attaches GPI anchors to proteins as they enter the lumen of the endoplasmic reticulum, is a membrane-bound hetero-pentameric complex consisting of Gpi8, Gpi16, Gaa1, Gpi17 and Gab1. Here, we expressed and purified the luminal domain of Saccharomyces cerevisiae (S. cerevisiae) Gpi8 using different expression systems, and examined its interaction with insect cell expressed luminal domain of S. cerevisiae Gpi16. We found that the N-terminal caspase-like domain of Gpi8 forms a disulfide-linked dimer, which is strengthened by N-glycosylation. The non-core domain of Gpi8 following the caspase-like domain inhibits this dimerization. In contrast to the previously reported disulfide linkage between Gpi8 and Gpi16 in human and trypanosome GPIT, our data show that the luminal domains of S. cerevisiae Gpi8 and S. cerevisiae Gpi16 do not interact directly, nor do they form a disulfide bond in the intact S. cerevisiae GPIT. Our data suggest that subunit interactions within the GPIT complex from different species may vary, a feature that should be taken into account in future structural and functional studies.
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GPI-T is a pentameric enzyme complex of unknown structure.
“Glycosylphosphatidylinositol transamidase (GPI-T) is a pentameric enzyme complex of unknown structure (88–90)”
From Computed structures of core eukaryotic protein complexes (2021) · cites this paper as [90]
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- Authors
- Yi, L., Bozkurt, G., Li, Q., Lo, S., Menon, A. K., & Wu, H.
- Affiliations
- Boston Children's Hospital (US)
- Weill Cornell Medicine (US)
- Corresponding author
- Wu, Hao
- Venue
- Scientific Reports
- Published
- 2017-04-04
- Volume
- 7
- Issue
- 1
- Pages
- 45912-45912
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC5379210
- OpenAlex
- W2605375582
- Cited by
- 12
- References
- 30
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2022: 4 · 2021: 2 · 2020: 1 · 2018: 1 · 2017: 1
- FWCI
- 0.34
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- eScholarship (California Digital Library) (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion, cc-by)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- disulfide bond formation, N-glycosylation, protein-protein interaction, disulfide-linked dimer, luminal domain, Saccharomyces cerevisiae, endoplasmic reticulum
- MeSH terms
- Protein Domains, Acyltransferases, Animals, Disulfides, Glycosylation, Humans, Insecta, Membrane Glycoproteins, Trypanosoma, Cell Adhesion Molecules, Glycosylphosphatidylinositols, Aminoacyltransferases, Saccharomyces cerevisiae Proteins, Protein Interaction Maps
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