用小分子抑制剂靶向S100钙结合蛋白
Targeting S100 Calcium-Binding Proteins with Small Molecule Inhibitors.
作者信息
Wilder Paul T, Varney Kristen M, Weber David J
机构信息
Center for Biomolecular Therapeutics (CBT), Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, USA.
University of Maryland Marlene and Stewart Greenebaum Comprehesive Cancer Center, Baltimore, MD, USA.
出版信息
Methods Mol Biol. 2019;1929:291-310. doi: 10.1007/978-1-4939-9030-6_19.
S100B is a small, dimeric, calcium-binding protein that is implicated in various diseases, most significantly cancer; therefore, there is interest in identifying S100B inhibitors that may have therapeutic value (Bresnick et al. Nat Rev Cancer 15:96-109, 2015; Chong et al. Curr Med Chem 23:1571-1596). Two fluorescence polarization competition assays (FPCA) are described here for S100B and S100A1 that are amenable to high-throughput screening (HTS) campaigns and can be used to determine the binding affinity (K ) of the inhibitors. One FPCA is used to identify and characterize inhibitors of S100B with the aim of finding new therapeutics, and the other was developed as a counter-screen to avoid inhibitors of S100A1 due to its role in regulating skeletal and cardiac muscle function. Also outlined are methods for expressing and purifying S100B and S100A1 in quantities needed for performing large HTS campaigns.
S100B是一种小型二聚体钙结合蛋白，与多种疾病有关，尤其是癌症；因此，人们对鉴定可能具有治疗价值的S100B抑制剂很感兴趣（布雷斯尼克等人，《自然评论：癌症》15:96 - 109，2015年；崇等人，《当代药物化学》23:1571 - 1596）。本文描述了两种适用于高通量筛选（HTS）活动的针对S100B和S100A1的荧光偏振竞争测定法（FPCA），可用于确定抑制剂的结合亲和力（K）。一种FPCA用于鉴定和表征S100B抑制剂，旨在寻找新的治疗方法，另一种则作为反筛选方法，以避免因S100A1在调节骨骼肌和心肌功能中的作用而导致的抑制剂干扰。文中还概述了在进行大规模HTS活动所需数量下表达和纯化S100B和S100A1的方法。
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