title: The effect of alpha-ketoglutaric acid on tyrosinase activity and conformation: Kinetics and molecular dynamics simulation study.
abstract: Alpha-ketoglutaric acid (AKG) is naturally found in organisms and is a well-known intermediate in the production of ATP or GTP in the Krebs cycle. We elucidated the effects of AKG on tyrosinase activity and conformation via methods of inhibition kinetics integrated with molecular dynamics (MD) simulations. AKG was found to be a reversible inhibitor of tyrosinase (IC50=15±0.5mM) and induced parabolic slope mixed-type inhibition. Based on our newly established equation, the dissociation constant (Kislope) was determined to be 7.93±0.31mM. The spectrofluorimetry studies showed that AKG mainly induced regional changes in the active site of tyrosinase, which reflects the flexibility of the active site. The computational docking and molecular dynamics (MD) simulations further demonstrated that AKG could interact with several residues near the substrate-binding site located in the tyrosinase active site pocket. Our study provides insight into the mechanism by which energy-producing intermediates such as AKG inhibit tyrosinase through its ketone groups. Also, AKG could be a potential natural antipigmentation agent due to its non-toxic property.
authors.affiliation: College of Biology and Science, Sichuan Agriculture University, Chengdu, 611130, PR China.
authors.forename: Lin
authors.initials: L
authors.lastname: Gou
authors.affiliation: Korean Bioinformation Center (KOBIC), Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, South Korea; Department of Nanobiotechnology and Bioinformatics, University of Sciences and Technology, Daejeon, 305-350, South Korea.
authors.forename: Jinhyuk
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Dermatology, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul, 135-710, South Korea.
authors.forename: Jun-Mo
authors.initials: JM
authors.lastname: Yang
authors.affiliation: Zhejiang Provincial Key Laboratory of Applied Enzymology, Yangtze Delta Region Institute of Tsinghua University, Jiaxing, 314006, PR China.
authors.forename: Yong-Doo
authors.initials: YD
authors.lastname: Park
authors.affiliation: Zhejiang Provincial Key Laboratory of Applied Enzymology, Yangtze Delta Region Institute of Tsinghua University, Jiaxing, 314006, PR China.
authors.forename: Hai-Meng
authors.initials: HM
authors.lastname: Zhou
authors.affiliation: College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, 310018, PR China. Electronic address: zhanyi_1006@hotmail.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: Zhejiang Provincial Key Laboratory of Applied Enzymology, Yangtze Delta Region Institute of Tsinghua University, Jiaxing, 314006, PR China. Electronic address: lzr7899@163.com.
authors.forename: Zhi-Rong
authors.initials: ZR
authors.lastname: Lü
mesh_terms: D020134:Catalytic Domain / Q000187:drug effects; D007656:Ketoglutaric Acids / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D014442:Monophenol Monooxygenase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D007656:Ketoglutaric Acids; D014442:Monophenol Monooxygenase
keywords: Alpha-ketoglutaric acid; Inhibition; Tyrosinase