Identification and Characterization of Proteins That Are Involved in RTP1S-Dependent Transport of Olfactory Receptors
Abstract
:1. Introduction
2. Results
2.1. Design and Construction of RTP1S and OR Fused with Biotin Ligase
2.2. Proximity Labeling-Coupled Mass Spectrometry in HEK293T Cells
2.3. HSPA6 and STAU2 Facilitate Surface Expression and Ligand Response of Olfr544
2.4. Intracellular Interaction Analysis Using NanoBiT Assay
3. Discussion
4. Materials and Methods
4.1. DNA and Vector Preparation
4.2. Cell Culture
4.3. Luciferase Assay
4.4. Affinity Purification of Biotinylated Proteins
4.5. MS Sample Preparation
4.6. LC-MS/MS Analysis
4.7. Data Analysis
4.8. Flow Cytometry Analyses
4.9. Split Luciferase Reconstruction Assay
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Inoue, R.; Fukutani, Y.; Niwa, T.; Matsunami, H.; Yohda, M. Identification and Characterization of Proteins That Are Involved in RTP1S-Dependent Transport of Olfactory Receptors. Int. J. Mol. Sci. 2023, 24, 7829. https://doi.org/10.3390/ijms24097829
Inoue R, Fukutani Y, Niwa T, Matsunami H, Yohda M. Identification and Characterization of Proteins That Are Involved in RTP1S-Dependent Transport of Olfactory Receptors. International Journal of Molecular Sciences. 2023; 24(9):7829. https://doi.org/10.3390/ijms24097829
Chicago/Turabian StyleInoue, Ryosuke, Yosuke Fukutani, Tatsuya Niwa, Hiroaki Matsunami, and Masafumi Yohda. 2023. "Identification and Characterization of Proteins That Are Involved in RTP1S-Dependent Transport of Olfactory Receptors" International Journal of Molecular Sciences 24, no. 9: 7829. https://doi.org/10.3390/ijms24097829