N-Glycan on the Non-Consensus N-X-C Glycosylation Site Impacts Activity, Stability, and Localization of the Sda Synthase B4GALNT2
Abstract
:1. Introduction
2. Results
2.1. Vertebrate B4GALNT2 Sequences Contain Unusual and Conserved N-Glycosylation Site(s)
2.2. N-Glycosylation of B4GALNT2 in Mammals
2.3. Impact of N-Glycan on B4GALNT2 Activity and Sda Synthesis
2.4. Impact of N-Glycan on Dimerization of B4GALNT2 Isoforms
2.5. Subcellular Localization of B4GALNT2 Isoforms
2.6. Model of the B4GALNT2 Structure
3. Discussion
4. Materials and Methods
4.1. Bioinformatic Analysis of B4GALNT Protein Sequences
4.2. Antibodies and Reagents
4.3. Plasmid Constructions of Human Long and Short B4GALNT2 Isoforms and Mutants
4.4. Cell Culture and Transfections
4.5. Western Blot Analysis
4.6. Total Proteins Preparation from Various Mammalian Gastrointestinal Tissues and Immunochemical Analyses
4.7. PNGase F and Endo H Treatments
4.8. B4GALNT2 Enzyme Activity
4.9. Flexibility Prediction
4.10. Modeling of the B4GALNT2 Dimeric Structure
- --is_prokaryote_list=false
- --max_template_date=2021-11-17
- --model_preset=multimer
- --db_preset=full_dbs
4.11. Statistical Methodology
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cogez, V.; Vicogne, D.; Schulz, C.; Portier, L.; Venturi, G.; de Ruyck, J.; Decloquement, M.; Lensink, M.F.; Brysbaert, G.; Dall’Olio, F.; et al. N-Glycan on the Non-Consensus N-X-C Glycosylation Site Impacts Activity, Stability, and Localization of the Sda Synthase B4GALNT2. Int. J. Mol. Sci. 2023, 24, 4139. https://doi.org/10.3390/ijms24044139
Cogez V, Vicogne D, Schulz C, Portier L, Venturi G, de Ruyck J, Decloquement M, Lensink MF, Brysbaert G, Dall’Olio F, et al. N-Glycan on the Non-Consensus N-X-C Glycosylation Site Impacts Activity, Stability, and Localization of the Sda Synthase B4GALNT2. International Journal of Molecular Sciences. 2023; 24(4):4139. https://doi.org/10.3390/ijms24044139
Chicago/Turabian StyleCogez, Virginie, Dorothée Vicogne, Céline Schulz, Lucie Portier, Giulia Venturi, Jérôme de Ruyck, Mathieu Decloquement, Marc F. Lensink, Guillaume Brysbaert, Fabio Dall’Olio, and et al. 2023. "N-Glycan on the Non-Consensus N-X-C Glycosylation Site Impacts Activity, Stability, and Localization of the Sda Synthase B4GALNT2" International Journal of Molecular Sciences 24, no. 4: 4139. https://doi.org/10.3390/ijms24044139