On selectivity and sensitivity of synthetic multifunctional pores as enzyme sensors: discrimination between ATP and ADP and comparison with biological pores.

This report delineates scope and limitation of the selectivity of synthetic multifunctional pores as enzyme sensors using glycolytic enzymes as example (G. Das, P. Talukdar, and S. Matile, Science, 2002, Vol. 298, pp. 1600-1602). Unproblematic detectability of hexokinase and phosphofructokinase demonstrates that the selectivity of synthetic multifunctional pore (SMPs) sensors suffices to sense ATP in mixed analytes containing ADP, whereas detection of the isomerization of glucose 6-phosphate into fructose 6-phosphate by phosphoglucose isomerase is not possible with confidence. The sensitivity of SMP sensors is sufficient for end-point detection of one picomole poly-L-glutamate hydrolyzed by papain in unoptimized assay format; the sensitivity of melittin as representative biological pore of similar charge and aggregation number to detect the same reaction is more than four orders of magnitude inferior.

[1]  Stefan Matile,et al.  Self-Assembled Rigid-Rod Ionophores , 1999 .

[2]  Stefan Matile,et al.  Rigid‐Rod β‐Barrel Ion Channels with Internal “Cascade Blue” Cofactors − Catalysis of Amide, Carbonate, and Ester Hydrolysis , 2002 .

[3]  S. Matile.  En route to supramolecular functional plasticity: artificial β-barrels, the barrel-stave motif, and related approaches , 2001 .

[4]  S. Matile,et al.  Giant Artificial Ion Channels Formed by Self-Assembled, Cationic Rigid-Rodβ-Barrels , 2000 .

[5]  L. Fothergill-Gilmore.  The evolution of the glycolytic pathway , 1986 .

[6]  S. Matile.  Bioorganic chemistry à la baguette: studies on molecular recognition in biological systems using rigid-rod molecules. , 2001, Chemical record.

[7]  B. Hess,et al.  Design of glycolysis. , 1981, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[8]  C. D. Hall,et al.  Supramolecular assemblies in natural and artificial ion channels , 2000 .

[9]  Stefan Matile,et al.  Synthetic catalytic pores. , 2003, Journal of the American Chemical Society.

[10]  Gopal Das,et al.  Enzyme screening with synthetic multifunctional pores: Focus on biopolymers , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[11]  P. Scrimin,et al.  Model membranes: developments in functional micelles and vesicles. , 1999, Current opinion in chemical biology.

[12]  S. Matile,et al.  α-Helix recognition by rigid-rod β-barrel ion channels with internal arginine-histidine dyads in polarized bilayer membranes , 2002 .

[13]  S. Matile,et al.  Recognition of polarized lipid bilayers by p-oligophenyl ion channels: from push-pull rods to push-pull barrels. , 2002, Journal of the American Chemical Society.

[14]  H. Bayley,et al.  Stochastic sensors inspired by biology , 2001, Nature.

[15]  S. Matile,et al.  On the electrostatics of cell-membrane recognition: from natural antibiotics to rigid push-pull rods. , 2000, Chemistry.

[16]  M. Adrian,et al.  β‐Fibrillogenesis from Rigid‐Rod β‐Barrels: Hierarchical Preorganization Beyond Microns , 2001 .

[17]  S. Matile,et al.  Synthetic multifunctional pores: lessons from rigid-rod β-barrels , 2003 .

[18]  O. Antunes,et al.  The High Resolution Crystal Structure of Yeast Hexokinase PII with the Correct Primary Sequence Provides New Insights into Its Mechanism of Action* , 2000, The Journal of Biological Chemistry.

[19]  George W. Gokel,et al.  Synthetic models of cation-conducting channels , 2001 .

[20]  S. Matile,et al.  On the Importance of Intermediate Internal Charge Repulsion for the Synthesis of Multifunctional Pores , 2002 .

[21]  Stefan Matile,et al.  p-Octiphenyl β-Barrels with Ion Channel and Esterase Activity , 2001 .

[22]  S. Matile,et al.  Voltage-dependent formation of anion channels by synthetic rigid-rod push-pull beta-barrels. , 2003, Chemistry.

[23]  S. Matile,et al.  Binding of Organic Anions by Synthetic Supramolecular Metallopores with Internal Mg2+–Aspartate Complexes , 2002, Chembiochem : a European journal of chemical biology.

[24]  Stefan Matile,et al.  Complementary characteristics of homologous p-octiphenyl beta-barrels with ion channel and esterase activity. , 2003, Bioorganic & medicinal chemistry.

[25]  Gopal Das,et al.  Fluorometric Detection of Enzyme Activity with Synthetic Supramolecular Pores , 2002, Science.

[26]  S. Matile,et al.  Transmembrane pores formed by synthetic p-octiphenyl β-barrels with internal carboxylate clusters: Regulation of ion transport by pH and Mg2+- complexed 8-aminonaphthalene-1,3,6-trisulfonate , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[27]  J. Ross,et al.  Oscillations and control features in glycolysis: numerical analysis of a comprehensive model. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[28]  S. Matile,et al.  Toward p-octiphenyl β-barrel RNases , 2002 .

[29]  Karen N. Allen,et al.  The Pentacovalent Phosphorus Intermediate of a Phosphoryl Transfer Reaction , 2003, Science.

[30]  Anthony W. Czarnik,et al.  Chemical Communication in Water Using Fluorescent Chemosensors , 1994 .

[31]  W. G. Miller.  Degradation of Poly-α,L-glutamic Acid. I. Degradation of High Molecular Weight PGA by Papain , 1961 .

[32]  S. Matile,et al.  Transmembrane B‐DNA , 2000, Chembiochem : a European journal of chemical biology.

[33]  W. Zygmunt.  Inhibition of Antibiotic Formation by Bromthymol Blue and Other Indicators in Streptomyces rimosus , 1962, Nature.

[34]  J. Reymond,et al.  Novel methods for biocatalyst screening. , 2001, Current opinion in chemical biology.

[35]  S. Matile,et al.  Synthetic multifunctional pores: deletion and inversion of anion/cation selectivity using pM and pH. , 2003, Organic & biomolecular chemistry.