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Shigehisa, R.

Publications and source records attributed to Shigehisa, R..

2 recordsLinked to original sources

Calibrated ADAMTS13 inhibition prevents cardiovascular shear coagulopathy

Mechanical circulatory support essential for managing severe heart failure frequently triggers bleeding complications, driven by shear stress-induced over-proteolysis of von Willebrand factor (VWF) by ADAMTS13. Inhibiting ADAMTS13 presents a rationale to treat this condition, known as acquired von Willebrand syndrome (AVWS). However, conventional therapeutic strategies remain limited due to the risk of triggering thrombotic thrombocytopenic purpura. Here we show that HA10, a humanized anti-ADAMTS13 antibody, preserves a residual level of ADAMTS13 activity above the thrombosis-associated threshold. Multimodal structural and biophysical analyses--including NMR, SAXS, and SANS--revealed that HA10 bound to the disintegrin-like domain of ADAMTS13, dynamically competing with VWF while leaving 10- 20% residual enzymatic activity. The therapeutic efficacy and safety of HA10 were verified in non-human primate models of AVWS. Our findings establish a novel paradigm of enzymatic calibration rather than complete blockade, offering a mechanistically targeted and safe therapeutic approach for cardiovascular bleeding.

biochemistry↗

A cyclo-DOPA 6-O-glucosyltransferase-mediated route for gomphrenin I biosynthesis in Basella alba and Gomphrena globosa

O_LIBetacyanins are red pigments characteristic of Caryophyllales and show considerable structural diversity, yet the enzymatic basis underlying 6-O-glucosylated betacyanins such as gomphrenin I has remained unclear. In particular, how alternative glucosylation patterns contribute to betacyanin diversification is poorly understood. C_LIO_LIHere, we identified cyclo-DOPA glucosyltransferases from Basella alba and Gomphrena globosa and examined their roles in gomphrenin I biosynthesis using transient expression assays and tobacco BY-2 cell systems. Phylogenetic analyses, structural modelling and site-directed mutagenesis were employed to investigate their functional and structural characteristics. C_LIO_LIBacDOPA5/6GTs catalysed both 5-O- and 6-O-glucosylation of cyclo-DOPA, leading to the production of betanin and gomphrenin I, whereas GgcDOPA6GT specifically mediated gomphrenin I formation. These enzymes belong to distinct subclades within the cDOPA-GT family, and mutational analyses demonstrated essential roles for conserved histidine residues and an -helical region adjacent to the catalytic site. C_LIO_LIThermal stability analyses further showed that gomphrenin I is more thermally stable than betanin, likely due to the formation of an intramolecular hydrogen bond. Together, these results reveal an additional cDOPA6GT-mediated route for gomphrenin I biosynthesis and provide insight into the diversification and functional specialization of betacyanins, linking the position of glucosylation to pigment stability and biochemical properties. C_LI

plant biology↗