title: Interactome analysis of AMP-activated protein kinase (AMPK)-α1 and -β1 in INS-1 pancreatic beta-cells by affinity purification-mass spectrometry.
abstract: The heterotrimeric enzyme AMP-activated protein kinase (AMPK) is a major metabolic factor that regulates the homeostasis of cellular energy. In particular, AMPK mediates the insulin resistance that is associated with type 2 diabetes. Generally, cellular processes require tight regulation of protein kinases, which is effected through their formation of complex with other proteins and substrates. Despite their critical function in regulation and pathogenesis, there are limited data on the interaction of protein kinases. To identify proteins that interact with AMPK, we performed large-scale affinity purification (AP)-mass spectrometry (MS) of the AMPK-α1 and -β1 subunits. Through a comprehensive analysis, using a combination of immunoprecipitaion and ion trap mass spectrometry, we identified 381 unique proteins in the AMPKα/β interactomes: 325 partners of AMPK-α1 and 243 for AMPK-β1. Further, we identified 196 novel protein-protein interactions with AMPK-α1 and AMPK-β1. Notably, in our bioinformatics analysis, the novel interaction partners mediated functions that are related to the regulation of actin organization. Specifically, several such proteins were linked to pancreatic beta-cell functions, including glucose-stimulated insulin secretion, beta-cell development, beta-cell differentiation, and cell-cell communication.
authors.affiliation: 1] Department of Biomedical Sciences, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [2] Department of Biomedical Engineering, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [3].
authors.forename: Sungyoon
authors.initials: S
authors.lastname: Moon
authors.affiliation: 1] Department of Biomedical Sciences, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [2] Department of Biomedical Engineering, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [3] Institute of Medical & Biological Engineering, Medical Research Center, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [4].
authors.forename: Dohyun
authors.initials: D
authors.lastname: Han
authors.affiliation: 1] Department of Biomedical Sciences, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [2] Department of Biomedical Engineering, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea.
authors.forename: Yikwon
authors.initials: Y
authors.lastname: Kim
authors.affiliation: 1] Department of Biomedical Sciences, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [2] Department of Biomedical Engineering, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea.
authors.forename: Jonghwa
authors.initials: J
authors.lastname: Jin
authors.affiliation: 1] Department of Biomedical Sciences, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [2] Department of Physilogy, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea.
authors.forename: Won-Kyung
authors.initials: WK
authors.lastname: Ho
authors.affiliation: 1] Department of Biomedical Sciences, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [2] Department of Biomedical Engineering, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea [3] Institute of Medical & Biological Engineering, Medical Research Center, Seoul National University College of Medicine, 28 Yongon-Dong, Seoul 110-799 Korea.
authors.forename: Youngsoo
authors.initials: Y
authors.lastname: Kim
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D000199:Actins / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D002846:Chromatography, Affinity; D019295:Computational Biology / Q000379:methods; D006801:Humans; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D013058:Mass Spectrometry; D058977:Molecular Sequence Annotation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D021122:Protein Subunits / Q000302:isolation & purification* / Q000378:metabolism*; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000199:Actins; D002352:Carrier Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D055372:AMP-Activated Protein Kinases