{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Eyster C"],"funding":["Atomwise AIMS Program","NHLBI NIH HHS","National Heart, Lung, and Blood Institute","National Institute of General Medical Sciences","NIGMS NIH HHS"],"pagination":["e0317167"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12097583"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["20(5)"],"pubmed_abstract":["The 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase (PFKFB) family of proteins are bifunctional enzymes that are of clinical relevance because of their roles in regulating glycolysis in insulin sensitive tissues and cancer. Here, we sought to express recombinant PFKFB2 and develop a robust protocol to measure its kinase activity. These studies resulted in the unexpected finding that bacterially expressed PFKFB2 is phosphorylated in situ on Ser483 but is not a result of autophosphorylation. Recombinant PFKFB2 was used to develop an enzymatic assay to test a library of molecules selected by the Atomwise AtomNet® AI platform. This resulted in the identification of a new inhibitor, B2, that inhibits PFKFB2 (IC50 3.29 μM) and PFKFB3 (IC50 11.89 μM). A-498 cells, which express both PFKFB2 an"],"journal":["PloS one"],"pubmed_title":["Mechanistic studies of PFKFB2 reveal a novel inhibitor of its kinase activity."],"pmcid":["PMC12097583"],"funding_grant_id":["A18-142","P20 GM139763","R01HL160955","P20GM139763","R01 HL160955"],"pubmed_authors":["Matsuzaki S","Faakye A","Humphries KM","Eyster C","Giorgione JR","Pranay A","Ahmed M"],"additional_accession":[]},"is_claimable":false,"name":"Mechanistic studies of PFKFB2 reveal a novel inhibitor of its kinase activity.","description":"The 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase (PFKFB) family of proteins are bifunctional enzymes that are of clinical relevance because of their roles in regulating glycolysis in insulin sensitive tissues and cancer. Here, we sought to express recombinant PFKFB2 and develop a robust protocol to measure its kinase activity. These studies resulted in the unexpected finding that bacterially expressed PFKFB2 is phosphorylated in situ on Ser483 but is not a result of autophosphorylation. Recombinant PFKFB2 was used to develop an enzymatic assay to test a library of molecules selected by the Atomwise AtomNet® AI platform. This resulted in the identification of a new inhibitor, B2, that inhibits PFKFB2 (IC50 3.29 μM) and PFKFB3 (IC50 11.89 μM). A-498 cells, which express both PFKFB2 an","dates":{"release":"2025-01-01T00:00:00Z","publication":"2025","modification":"2026-06-02T20:59:11.33Z","creation":"2026-04-20T03:14:29.69Z"},"accession":"S-EPMC12097583","cross_references":{"pubmed":["40402947"],"doi":["10.1371/journal.pone.0317167"]}}