Enhancing catalytic activity of multi-enzyme in vitro through substrate channeling effect is promis-ing yet challenging.Herein,conjugated microporous polymers(CMPs)-scaffolded integrated en-zyme cascade systems(I-ECSs...Enhancing catalytic activity of multi-enzyme in vitro through substrate channeling effect is promis-ing yet challenging.Herein,conjugated microporous polymers(CMPs)-scaffolded integrated en-zyme cascade systems(I-ECSs)are constructed through co-entrapping glucose oxidase(GOx)and horseradish peroxidase(HRP),in which hydrogen peroxide(H_(2)O_(2)) is the intermediate product.The interplay of low-resistance mass transfer pathway and appropriate pore wall-H_(2)O_(2) interactions facilitates the directed transfer of H_(2)O_(2),resulting in 2.4-fold and 5.0-fold elevation in catalytic activ-ity compared to free ECSs and separated ECSs,respectively.The substrate channeling effect could be regulated by altering the mass ratio of GOx to HRP.Besides,I-ECSs demonstrate excellent stabili-ties in harsh environments and multiple recycling.展开更多
As a type II or III transmembrane glycoprotein, human CD38 is ubiquitously expressed in all mammalian tissues. CD38 is a multi-functional enzyme and a member of the ADP-ribosyl cyclase family, and it catalyzes nicotin...As a type II or III transmembrane glycoprotein, human CD38 is ubiquitously expressed in all mammalian tissues. CD38 is a multi-functional enzyme and a member of the ADP-ribosyl cyclase family, and it catalyzes nicotinamide adenine dinucleotide （NAD^＋） and nicotinamide adenine dinucleotide phosphate （NADP＋） to two distinct Ca^2＋ messengers as follows： cyclic ADP-ribose （cADPR） and nicotinic acid adenine dinucleotide phosphate （NAADP）, respectively. Moreover, both cADPR and NAADP mediate mobilization of intracellular Ca^2＋ targeting endoplasmic stores and the lysosomes, respectively. In this study, we combined ligand-based and structure-based virtual screening strategies to compare the inhibitor discovery efficacy based on natural substrates and the known inhibitors. The similarity queries towards SPECS database were carried out using ROCS and EON modules of OpenEye software. The hits were further docked to CD38 using AutoDock 4.05 program. In addition, ADME studies were also processed considering solubility in water and membrane permeability. Finally, we identified 17 compotmds-based on natural substrates and 10 compounds based on known inhibitor models. The results showed that the known inhibitor H2-based model was more efficient in virtual screening of CD38 non-covalent inhibitors.展开更多
文摘Enhancing catalytic activity of multi-enzyme in vitro through substrate channeling effect is promis-ing yet challenging.Herein,conjugated microporous polymers(CMPs)-scaffolded integrated en-zyme cascade systems(I-ECSs)are constructed through co-entrapping glucose oxidase(GOx)and horseradish peroxidase(HRP),in which hydrogen peroxide(H_(2)O_(2)) is the intermediate product.The interplay of low-resistance mass transfer pathway and appropriate pore wall-H_(2)O_(2) interactions facilitates the directed transfer of H_(2)O_(2),resulting in 2.4-fold and 5.0-fold elevation in catalytic activ-ity compared to free ECSs and separated ECSs,respectively.The substrate channeling effect could be regulated by altering the mass ratio of GOx to HRP.Besides,I-ECSs demonstrate excellent stabili-ties in harsh environments and multiple recycling.
基金National Natural Science Foundation of China(Grant No.21272017 and 81172917)
文摘As a type II or III transmembrane glycoprotein, human CD38 is ubiquitously expressed in all mammalian tissues. CD38 is a multi-functional enzyme and a member of the ADP-ribosyl cyclase family, and it catalyzes nicotinamide adenine dinucleotide （NAD^＋） and nicotinamide adenine dinucleotide phosphate （NADP＋） to two distinct Ca^2＋ messengers as follows： cyclic ADP-ribose （cADPR） and nicotinic acid adenine dinucleotide phosphate （NAADP）, respectively. Moreover, both cADPR and NAADP mediate mobilization of intracellular Ca^2＋ targeting endoplasmic stores and the lysosomes, respectively. In this study, we combined ligand-based and structure-based virtual screening strategies to compare the inhibitor discovery efficacy based on natural substrates and the known inhibitors. The similarity queries towards SPECS database were carried out using ROCS and EON modules of OpenEye software. The hits were further docked to CD38 using AutoDock 4.05 program. In addition, ADME studies were also processed considering solubility in water and membrane permeability. Finally, we identified 17 compotmds-based on natural substrates and 10 compounds based on known inhibitor models. The results showed that the known inhibitor H2-based model was more efficient in virtual screening of CD38 non-covalent inhibitors.
