The Tetraspanin Tspan8 Associates with Endothelin Converting Enzyme ECE1 and Regulates Its Activity
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Cell Lines
2.2. Antibodies
2.3. Production of Tspan8 Knockout Mice
2.4. Endothelin Dosage
2.5. Preparation of Intestinal Tissues and DNA Dosage
2.6. Immunofluorescence
2.7. Immunoprecipitation (IP) and Biotinylation of Cell Surface Proteins
2.8. Mass Spectrometry Protein Identifications [15]
3. Results
3.1. Tspan8 Molecular Partners
3.2. Association of ECE1 with Tspan8 Is Confirmed by Western Blot
3.3. ECE1 Is Associated with Tspan8 at the Plasma Membrane
3.4. Modulation of bigET1 to Endothelin Conversion by Cellular ECE1 in the Presence of Tspan8
3.5. Tspan8 Regulation of BigET1 Conversion by Intestinal Tissue
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Proteins (Genes Names) | Isreco1—IP CD9 | Is1-tspan8—IP CD9 | Is1-tspan 8—IP Tspan8 | ||||||
---|---|---|---|---|---|---|---|---|---|
Peptides (n) | S-Area | Peptides (n) | S-Area | Peptides (n) | S-Area | ||||
Total | Unique | Total | Unique | Total | Unique | ||||
ECE1 | 0 | 0 | 0 | 10 | 3 | 2.62 × 107 | 15 | 7 | 1.13 × 108 |
Tspan6 | 0 | 0 | 0 | 3 | 1 | 3.64 × 107 | 4 | 2 | 2.56 × 107 |
ITGB1 | 29 | 18 | 2.72 × 109 | 28 | 17 | 3.03 × 109 | 27 | 16 | 2.43 × 109 |
NOTCH2 | 5 | 1 | 7.98 × 106 | 3 | 1 | 1.30 × 107 | 5 | 3 | 5.07 × 106 |
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Zhu, Y.; Saint-Pol, J.; Nguyen, V.; Rubinstein, E.; Boucheix, C.; Greco, C. The Tetraspanin Tspan8 Associates with Endothelin Converting Enzyme ECE1 and Regulates Its Activity. Cancers 2023, 15, 4751. https://doi.org/10.3390/cancers15194751
Zhu Y, Saint-Pol J, Nguyen V, Rubinstein E, Boucheix C, Greco C. The Tetraspanin Tspan8 Associates with Endothelin Converting Enzyme ECE1 and Regulates Its Activity. Cancers. 2023; 15(19):4751. https://doi.org/10.3390/cancers15194751
Chicago/Turabian StyleZhu, Yingying, Julien Saint-Pol, Viet Nguyen, Eric Rubinstein, Claude Boucheix, and Céline Greco. 2023. "The Tetraspanin Tspan8 Associates with Endothelin Converting Enzyme ECE1 and Regulates Its Activity" Cancers 15, no. 19: 4751. https://doi.org/10.3390/cancers15194751