title: FK506, an Immunosuppressive Drug, Induces Autophagy by Binding to the V-ATPase Catalytic Subunit A in Neuronal Cells.
abstract: The drug FK506 (tacrolimus, fujimycin) exerts its immunosuppressive effects by regulating the nuclear factor of the activated T-cell (NFAT) family of transcription factors. However, FK506 also exhibits neuroprotective effects, but its direct target proteins that mediate these effects have not been determined. To identify the target proteins responsible for FK506's neuroprotective effects, the drug affinity responsive target stability (DARTS) method was performed using label-free FK506, and LC-MS/MS analysis of the FK506-treated proteome was also performed. Using DARTS and LC-MS/MS analyses in combination with reference studies, V-ATPase catalytic subunit A (ATP6V1A) was identified as a new target protein of FK506. The biological relevance of ATP6V1A in mediating the neuroprotective effects of FK506 was validated by analyzing FK506 activity with respect to autophagy via acridine orange staining and transcription factor EB (TFEB) translocation assay. These analyses demonstrated that the binding of FK506 with ATP6V1A induces autophagy by activating the translocation of TFEB from the cytosol into the nucleus. Because autophagy has been identified as a mechanism for treating neurodegenerative diseases and because we have demonstrated that FK506 induces autophagy, this study demonstrates that FK506 is a possible new therapy for treating neurodegenerative diseases.
authors.affiliation: Global Research Laboratory, Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University , Seoul 120-749, Korea.
authors.forename: Dongyoung
authors.initials: D
authors.lastname: Kim
authors.affiliation: Global Research Laboratory, Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University , Seoul 120-749, Korea.
authors.forename: Hui-Yun
authors.initials: HY
authors.lastname: Hwang
authors.affiliation: Biomedical Omics Group, Korea Basic Science Institute , Ochang, Chungbuk 28119, Korea.
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Biomedical Omics Group, Korea Basic Science Institute , Ochang, Chungbuk 28119, Korea.
authors.forename: Ju Yeon
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Biomedical Omics Group, Korea Basic Science Institute , Ochang, Chungbuk 28119, Korea.
authors.forename: Jong Shin
authors.initials: JS
authors.lastname: Yoo
authors.affiliation: Clinical Protein Science & Imaging, Biomedical Center, Department of Biomedical Engineering, Lund University , BMC D13, SE-221 84 Lund, Sweden.
authors.forename: György
authors.initials: G
authors.lastname: Marko-Varga
authors.affiliation: Global Research Laboratory, Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University , Seoul 120-749, Korea.|Department of Internal Medicine, College of Medicine, Yonsei University , Seoul 120-752, Korea.
authors.forename: Ho Jeong
authors.identifier: 0000-0002-6919-833X
authors.initials: HJ
authors.lastname: Kwon
mesh_terms: D000165:Acridine Orange / Q000737:chemistry; D001343:Autophagy / Q000187:drug effects*; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005786:Gene Expression Regulation; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011485:Protein Binding; D021381:Protein Transport / Q000187:drug effects; D000071456:Sequestosome-1 Protein / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D016559:Tacrolimus / Q000494:pharmacology*; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics* / Q000378:metabolism
chemical_list: D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D007166:Immunosuppressive Agents; C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins; D018696:Neuroprotective Agents; C099718:SQSTM1 protein, human; D000071456:Sequestosome-1 Protein; C064972:TFEB protein, human; C581697:ATP6V1A protein, human; D025262:Vacuolar Proton-Translocating ATPases; D000165:Acridine Orange; D016559:Tacrolimus
keywords: DARTS; FK506; LC−MS/MS; TFEB translocation; V-ATPase; autophagy; neuroprotective activity