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. 2022 Mar 21;298(5):101853. doi: 10.1016/j.jbc.2022.101853

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Co-immunoprecipitation of DNPB enzymes with PAICS following anti-StrepTag pull-down from crPAICS::2×Strep-PAICS and crPAICS::PAICS-2×Strep cells.A, Western blots showing the co-IP of PFAS, GART, MTHFD1, and ATIC with StrepTag-labeled PAICS from rescued crPAICS cells cultured under both purine-rich (P+) and purine-depleted (P−) conditions. B, Western blots of endogenous PAICS co-immunoprecipitated by 2×Strep-PAICS or PAICS-2×Strep overexpressed in HEK293T; EGFP-2×Strep is used as the noninteracting co-IP control. ATIC, 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase; DNPB, de novo purine biosynthesis; GART, phosphoribosylglycinamide synthetase/formyltransferase/phosphoribosylaminoimidazole synthetase; PAICS, phosphoribosylaminoimidazole carboxylase/succinocarboxamide synthetase; PFAS, phosphoribosylformylglycinamidine synthase.