Design,synthesis, and evaluation of a selective chemosensor for leucine-rich repeat kinase 2 |
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| Affiliation: | 1. College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University, Seoul 151-742, Republic of Korea;2. New Drug Preclinical & Analytical Team, Life Science R&D Center, SK Chemicals, 310 Pangyo-ro, 463-400, Republic of Korea;3. College of Pharmacy and BK21 PLUS R-FIND Team, Dongguk University-Seoul, 32 Dongguk-lo, Ilsandong-gu, Goyang, Gyeonggi-do 410-820, Republic of Korea;4. College of Pharmacy and Integrated Research Institute of Pharmaceutical Sciences, The Catholic University of Korea, Bucheon 420-743, Republic of Korea;1. Department of Pharmaceutical Chemistry, Y.B. Chavan College of Pharmacy, Dr. Rafiq Zakaria Campus, Rauza Bagh, P.B. No. 33, Aurangabad 431001, M.S., India;2. Department of Chemical Technology, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad 431004, M.S., India;1. Department of Medicinal Chemistry, School of Pharmacy, Fudan University, 826 Zhangheng Rd, Shanghai 201203, PR China;2. Key Laboratory of Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Fudan University Shanghai Medical College, Shanghai 200032, PR China;3. State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Rd, Shanghai 201203, PR China;1. Department of Pharmaceutical Science, Università degli Studi, Via Celoria 2, 20100 Milano, Italy;2. Department of Health Sciences, Università degli Studi, Via A. Di Rudinì 8, 20142 Milano, Italy;3. S. Paolo Hospital, Via A. Di Rudinì 8, 20142 Milano, Italy;1. Institut de Pharmacologie de Sherbrooke, 3001, 12e av nord, Sherbrooke (Qc) J1H 5N4, Canada;2. SCF Pharma, Ste Luce (Qc) G0K 1P0, Canada |
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| Abstract: | We describe the design, synthesis, and evaluation of a selective activity probe for leucine-rich repeat kinase 2 (LRRK2), a possible molecular target for the treatment of Parkinson’s disease. Our optimal chemosensor design, termed Nictide-S2, incorporates a phosphorylation-sensitive sulfonamido-oxine fluorophore at an engineered cysteine within the substrate sequence. This design allows for the direct, real-time analysis of LRRK2 kinase activity with a detection limit of 2.5 nM. Under optimized conditions, we measured a Z′ factor of 0.7 demonstrating the potential utility of this assay for inhibitor screening. Off-target kinases capable of phosphorylating Nictide-S2 are identified and an optimized inhibitor cocktail for suppressing background signal is provided. The resulting chemosensor could be utilized to identify LRRK2 inhibitors as well as selectively report on LRRK2 activity in the presence of off-target kinases. |
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| Keywords: | Fluorescence-based biosensor Kinase activity assay LRRK2 Parkinson’s disease Inhibitor |
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