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An in-depth, detailed review of recent TrpS research describing mutational studies, novel transition-state analogs and the use of novel ASLs both in solution and in crystallographic studies. Points of focus include the dual role of βGlu109 as well as β-site dynamics and its regulation by ASL binding.
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The authors present a series of novel IGP analogs that bind to the α-site with high specificity and that mimic the allosteric behavior of IGP and G3P. Crystal structures of these compounds bound to the E(Ain) enzyme are described. As these IGP analogs induce the closed form of the α-site and stabilize the closed form of the β-site they represent powerful tools for the study of allosteric regulation in TrpS.
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Ngo H., Harris R., Kimmich N., Casino P., Niks D., Blumenstein L., Barends T., Kulik V., Weyand M., Schlichting I., et al. Synthesis and Characterization of allosteric probes of substrate channeling in the tryptophan synthase bienzyme complex. Biochemistry 46 (2007) 7713-7727. The authors present a series of novel IGP analogs that bind to the α-site with high specificity and that mimic the allosteric behavior of IGP and G3P. Crystal structures of these compounds bound to the E(Ain) enzyme are described. As these IGP analogs induce the closed form of the α-site and stabilize the closed form of the β-site they represent powerful tools for the study of allosteric regulation in TrpS.
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•] on Stage I of the β-reaction. The authors compare structures of the analog complexes/E(Ain) and E(AA) structures, and by adding their structural and kinetic findings to the existing data arrive at a detailed, step-by-step mechanism for TrpS catalysis and regulation.
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•] and the spectroscopic probe trans-3-indole-3′-acrylate that mimics l-Trp, the authors show that in response to α-site ligand binding the β-site switches between low-affinity and high-affinity conformations in the internal aldimine state.
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•] and the spectroscopic probe trans-3-indole-3′-acrylate that mimics l-Trp, the authors show that in response to α-site ligand binding the β-site switches between low-affinity and high-affinity conformations in the internal aldimine state.
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2) intermediate in the β-reaction were prepared in a TrpS crystal and the 2.1 Å resolution crystal structure was determined. The structure shows the roles of αGlu49 and αAsp60 in α-site catalysis. The indoline quinonoid represents the first high-quality structure of a PLP-enzyme quinonoid and confirms the role of βGlu109 in Stage II catalysis.
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