title: Fragment-based methods for the discovery of inhibitors modulating lysyl-tRNA synthetase and laminin receptor interaction.
abstract: Lysyl-tRNA synthetase (KRS) is an enzyme that conjugates lysine to its cognate tRNAs in the process of protein synthesis. In addition to its catalytic function, KRS binds to the 67-kDa laminin receptor (LR) on the cell membrane and facilitates cell migration and metastasis. Modulation of this interaction by small-molecule inhibitors can be exploited to suppress cancer metastasis. In this study, we present fragment-based methods for the identification of inhibitors and monitoring protein-protein interactions between KRS and LR. First, we identified the amino acid residues, located on the KRS anticodon-binding domain, which interact with the C-terminal extension of the LR. One-dimensional (1D) relaxation-edited nuclear magnetic resonance spectroscopy (NMR) and competition experiments were designed and optimized to screen the fragment library. For screening using two-dimensional (2D) NMR, we identified the indicative signals in the KRS anticodon-binding domain and selected inhibitors that bind to KRS and compete with LR at the KRS-LR binding interface. These methods may offer an efficient approach for the discovery of anti-metastatic drugs.
authors: College of Pharmacy, Korea University, Sejong 30019, Republic of Korea.
authors: Hye Young
authors: HY
authors: Cho
authors: Medicinal Bioconvergence Research Center, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: sungkim@snu.ac.kr.
authors: Sunghoon
authors: S
authors: Kim
authors: College of Pharmacy, Korea University, Sejong 30019, Republic of Korea. Electronic address: yhjeon@korea.ac.kr.
authors: Young Ho
authors: YH
authors: Jeon
mesh_terms: D020816:Amino Acid Motifs; D000926:Anticodon / Q000737:chemistry / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology; D001665:Binding Sites; D055808:Drug Discovery / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D057166:High-Throughput Screening Assays; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008250:Lysine-tRNA Ligase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D012357:RNA, Transfer, Lys / Q000235:genetics / Q000378:metabolism; D017474:Receptors, Laminin / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology*; D046249:Transfer RNA Aminoacylation
chemical_list: D000926:Anticodon; D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D012357:RNA, Transfer, Lys; D017474:Receptors, Laminin; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; D008250:Lysine-tRNA Ligase; D008239:Lysine
keywords: Cancer metastasis; Fragment-based drug discovery; Laminin receptor; Lysyl-tRNA synthetase; Nuclear magnetic resonance (NMR); Protein–protein interaction