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Pocket incoming11/13/2022 ![]() This path enables substrate-product assisted processive catalysis through multiple hydrolytic events without HvExoI losing contact with oligo- or polymeric substrates. Structural analyses and multi-scale molecular modelling of nanoscale reactant movements in HvExoI reveal that upon productive binding of incoming substrates, the glucose product modifies its binding patterns and evokes the formation of a transient lateral cavity, which serves as a conduit for glucose departure to allow for the next catalytic round. Here, we report that the alkyl β-D-glucoside and methyl 6-thio-β-gentiobioside substrate analogues perfused in crystalline HvExoI bind across the catalytic site after they displace glucose, while methyl 2-thio-β-sophoroside attaches nearby. The high-resolution structure of the native Hordeum exo-hydrolase HvExoI isolated from seedlings reveals that non-covalently trapped glucose forms a stable enzyme-product complex. associate and products dissociate from enzyme catalytic sites rapidly, which hampers investigations of their trajectories. 15 School of Life Sciences, Huaiyin Normal University, Huaian, 223300, China. 14 School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Glen Osmond South Australia, 5064, Australia.13 Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193, Spain.12 Institute of Chemical Research of Catalonia, The Barcelona Institute of Science and Technology, Tarragona, 43007, Spain.11 Departament de Química, Universitat Autònoma de Barcelona, Bellaterra, 08193, Spain.10 Departamento de Ciencias Químicas, Universidad Andrés Bello, Sede Concepción, Talcahuano, 4260000, Chile.9 Institució Catalana de Recerca i Estudis Avançats, Barcelona, 08010, Spain.8 Departament de Química Inorgànica i Orgànica, Universitat de Barcelona, Barcelona, 08028, Spain.7 Centre for Cooperative Research in Biosciences, Derio-Bizkaia, 48160, Spain.6 Institute of Chemistry, Slovak Academy of Sciences, Bratislava, 84538, Slovak Republic.5 GE Healthcare Life Sciences, Paramatta NSW, 2150, Australia.4 University Grenoble Alpes, Centre de Recherches sur les Macromolécules Végétales, Grenoble cedex 9, 38041, France.3 School of Chemistry and Center for Biomolecular Structure, Function and Application, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand.2 School of Agriculture, Food and Wine, University of Adelaide, Waite Campus, Glen Osmond South Australia, 5064, Australia.1 Commonwealth Scientific and Industrial Research Organisation, Materials Science and Engineering, Parkville Victoria, 3052, Australia. ![]()
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