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Miyaguchi, I.

Publications and source records attributed to Miyaguchi, I..

3 recordsLinked to original sources

Resolution-standardized evaluation of ligand atomic coordinates in crystallographic structures using machine learning

Accurate assessment of ligand coordinate-density consistency across different resolutions remains challenging in macromolecular crystallography. We introduce the atomic Box Correlation Coefficient (aBCC), an atom-level metric for evaluating the consistency between ligand atomic coordinates and electron density in a resolution-standardized framework. To predict aBCC values from electron-density maps, we developed QAEmap, a machine-learning model based on three-dimensional convolutional neural networks (3D-CNNs). The model was trained using Fourier-truncated electron-density maps and corresponding ligand coordinates generated from high-resolution structures in the Protein Data Bank. It was evaluated using both Fourier-truncated electron-density maps and experimentally determined PDB structures. was evaluated using both Fourier-truncated electron-density maps and experimentally determined PDB structures.The prediction accuracy gradually decreased with decreasing resolution, but remained reliable up to [~]3.5 [A]. These results demonstrate that aBCC enables resolution-standardized atom-wise evaluation of coordinate-density consistency across different resolutions and provide a foundation for further development and refinement of machine learning-based coordinate validation. SynopsisWe introduce the atomic box correlation coefficient (aBCC), a machine learning-based metric for the resolution-standardized atom-level evaluation of ligand coordinate-density consistency in crystallographic structures. aBCC provides a common framework for assessing and communicating the local coordinate reliability between structural biologists and researchers in structure-based drug discovery.

molecular biology↗

Structure-guided development of a potent human B0AT1 inhibitor effective in a mouse model of phenylketonuria

B0AT1 is a neutral amino acid transporter responsible for the (re)absorption in the intestine and kidney. Here, we developed small-molecule inhibitors of B0AT1 as a therapeutic strategy to a transient pocket, located [~]17 [A] away from the substrate-binding site and unique to the outward-open conformation, and stabilize an outward-occluded conformation that prevents the conformational transitions required for transport. Guided by structural insights, we optimized an initial inhibitor (Cinromide) to improve potency and cross-species activity, yielding compound 3, which inhibits both human and mouse B0AT1 with submicromolar IC_50 values. In a PKU mouse model (Pahenu2), oral administration of compound 3 increased urinary Phe excretion and significantly reduced plasma Phe. Our findings identify a druggable allosteric site in B0AT1, demonstrate its utility for achieving potent and selective inhibition in vivo, and establish allosteric blockade as a therapeutic approach for PKU and other SLC6-family transporters.

pharmacology and toxicology↗

Structural insights into the mechanism of the human SGLT2-MAP17 glucose transporter

Selective sodium-glucose cotransporter 2 (SGLT2) plays an important role in glucose reabsorption. SGLT2 inhibitors suppress glucose reabsorption from the kidneys, thus reducing blood glucose levels in type 2 diabetes patients. We and other groups have developed several SGLT2 inhibitors starting from a natural product, phlorizin, but their action mechanisms remain unknown. Here, we elucidated the physiological hSGLT2-MAP17 complex structures bound to five SGLT2 inhibitors using single-particle cryo-electron microscopy. Canagliflozin, dapagliflozin, TA-1887, and sotagliflozin were bound in the outward-facing structure, whereas phlorizin was bound in the inward-open structure. The phlorizin-hSGLT2 interaction biochemically exhibited biphasic binding. Phlorizin weakly binds, via the phloretin motif, from its intracellular side near the Na+-binding site, while strongly interacts from its extracellular side. Unexpectedly, bound Na+ stabilizes the outward-open conformation, while its release allows the transporter to adopt inward-open state. Our results first visualized the Na+-binding and inward-open conformation of hSGLT2-MAP17, clarifying the unprecedented Na+-dependent sugar transport mechanism with MAP17 acting as a scaffold, and may pave the way for development of next-generation SGLT inhibitors.

molecular biology↗