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Protein interactome of the deamidase phosphoribosylformylglycinamidine synthetase (PFAS) by LC-MS/MS
- Lu, Ai, Disoma, Cyrollah, Zhou, Yuzheng, Chen, Zongpeng, Zhang, Liming, Shen, Yilun, Zhou, Mei, Du, Ashuai, Zheng, Rong, Li, Sijia, Alsaadawe, Moyed, Li, Shiqin, Li, Jiada, Wang, Weilan, Jiang, Taijiao, Peng, Jian, Xia, Zanxian
- Biochemical and biophysical research communications 2019 v.513 no.3 pp. 746-752
- Western blotting, bioinformatics, carcinogenesis, deamidation, enzymes, gene ontology, liquid chromatography, magnetism, neoplasms, post-translational modification, precipitin tests, protein-protein interactions, proteins, tandem mass spectrometry
- Phosphoribosylformylglycinamidine synthase (PFAS) is an essential enzyme in de novo synthesis of purine. Previously, PFAS has been reported to modulate RIG-I activation during viral infection via deamidation. In this study, we sought to identify potential substrates that PFAS can deamidate. Flag-PFAS was transfected into HEK-293T cells and PFAS associated proteins were purified with anti-Flag M2 magnetic beads. PFAS associated proteins were identified using mass spectrometry and were analyzed using bioinformatics tools including KEGG pathway analysis, gene ontology annotation, and protein interaction network analysis. A total of 441 proteins is suggested to potentially interact with PFAS. Of this number, 12 were previously identified and 429 are newly identified. The interactions of PFAS with CAD, CCT2, PRDX1, and PHGDH were confirmed by co-immunoprecipitation and western blotting. This study is first to report the interaction of PFAS with several proteins which play physiological roles in tumor development including CAD, CCT2, PRDX1, and PHGDH. Furthermore, we show here that PFAS is able to deamidate PHGDH, and induce other posttranslational modification into CAD, CCT2 and PRDX1. The present data provide insight on the biological function of PFAS. Further study to explore the role of these protein interactions in tumorigenesis and other diseases is recommended.