Methods for electronic fluorescent perturbation for analysis and electronic perturbation catalysis for synthesis

Abstract

Methods for catalyzing cleavage of a bond with an electric field device. In one method, a catalytic peptide having a first reactive group and a second reactive group is coupled to an electrode. The catalytic peptide is then contacted with a solution containing a substrate. The first reactive group reacts with the substrate to form a first intermediate. The second reactive group then reacts with the first intermediate to form a positively-charged second intermediate having an acyl bond and a negatively-charged first reactive group. An electronic pulsing sequence is then applied to the electrode to separate the negatively-charged first reactive group and the positively-charged second intermediate. The second intermediate is then reacted by acyl transfer to cleave the acyl bond. The first reactive group may be a sulfhydryl or deprotonated sulfhydryl. The second reactive group may be an imidazole. The substrate may contain an ester or amide bond.

1 . A method for catalyzing a cleavage of a bond with an electric field device, comprising the steps of: 
 coupling a catalytic peptide sequence to an electrode, wherein the catalytic peptide sequence has a first reactive group and a second reactive group;    contacting the catalytic peptide sequence that is coupled to the electrode with a solution containing a substrate to be hydrolyzed;    reacting the first reactive group with the substrate to form a first intermediate;    reacting the second reactive group with the first intermediate to form a positively charged second intermediate and a negatively charged first reactive group, wherein the positively charged second intermediate comprises an acyl bond;    applying an electronic pulsing sequence to the electrode to separate the negatively charged first reactive group from the positively charged second intermediate; and    reacting the second intermediate by acyl transfer to cleave the acyl bond.    
     
     
         2 . The method of  claim 1 , wherein the first reactive group is a sulfhydryl.  
     
     
         3 . The method of  claim 1 , wherein the first reactive group is a deprotonated sulfhydryl group.  
     
     
         4 . The method of  claim 3 , wherein the deprotonated sulfhydryl group is formed from a de-protonated cysteine.  
     
     
         5 . The method of  claim 1 , wherein the second reactive group is an imidazole.  
     
     
         6 . The method of  claim 5 , wherein the imidazole is part of a histidine.  
     
     
         7 . The method of  claim 1 , wherein the acyl bond is an amide.  
     
     
         8 . The method of  claim 1 , wherein the catalytic peptide is coupled to a permeation layer associated with the electrode.  
     
     
         9 . The method of  claim 1 , wherein the substrate comprises an ester.  
     
     
         10 . The method of  claim 1 , wherein the substrate comprises an amide.  
     
     
         11 . The method of  claim 1 , wherein the step of reacting the second intermediate by acyl transfer to cleave the acyl bond comprises hydrolyzing the acyl bond.  
     
     
         12 . A method for catalyzing cleavage of a bond with an electric field device, comprising the steps of: 
 coupling a catalytic peptide sequence to an electrode, wherein the catalytic peptide sequence has a deprotonoated sulfhydryl group and an imidazole;    contacting the catalytic peptide that is coupled to the electrode with a solution containing a substrate to be hydrolyzed;    reacting the deprotonoated sulfhydryl group with the substrate to form a first intermediate;    reacting the imidazole with the first intermediate to form a positively charged second intermediate and a negatively charged deprotonoated sulfhydryl group, wherein the positively charged second intermediate comprises an acyl bond;    applying an electronic pulsing sequence to the electrode to separate the negatively charged deprotonoated sulfhydryl group from the positively charged second intermediate; and    reacting the second intermediate by acyl transfer to cleave the acyl bond.    
     
     
         13 . The method of  claim 12 , wherein the deprotonoated sulfhydryl group is formed from a de-protonated cysteine.  
     
     
         14 . The method of  claim 12 , wherein the imidazole is part of a histidine.  
     
     
         15 . The method of  claim 12 , wherein the acyl bond is an amide.  
     
     
         16 . The method of  claim 12 , wherein the catalytic peptide is coupled to a permeation layer associated with the electrode.  
     
     
         17 . The method of  claim 12 , wherein the step of reacting the second intermediate by acyl transfer to cleave the acyl bond comprises hydrolyzing the acyl bond.  
     
     
         18 . The method of  claim 12 , wherein the substrate comprises an ester.  
     
     
         19 . The method of  claim 12 , wherein the substrate comprises an amide.  
     
     
         20 . A method for catalyzing cleavage of a bond with an electric field device, comprising the steps of: 
 coupling a catalytic peptide sequence to an electrode, wherein the catalytic peptide sequence has a thiol and an imidazole;    contacting the catalytic peptide that is coupled to the electrode with a solution containing a substrate to be hydrolyzed;    reacting the thiol with the substrate to form a first intermediate;    reacting the imidazole with the first intermediate to form a positively charged second intermediate and a negatively charged deprotonated thiol, wherein the positively charged second intermediate comprises an acyl bond;    applying an electronic pulsing sequence to the electrode to separate the negatively charged deprotonated thiol from the positively charged second intermediate; and    reacting the second intermediate by acyl transfer to cleave the acyl bond.    
     
     
         21 . The method of  claim 20 , wherein the thiol is part of a cysteine.  
     
     
         22 . The method of  claim 20 , wherein the imidazole is part of a histidine.  
     
     
         23 . The method of  claim 20 , wherein the acyl bond is an amide.  
     
     
         24 . The method of  claim 20 , wherein the catalytic peptide is coupled to a permeation layer associated with the electrode.  
     
     
         25 . The method of  claim 20 , wherein the step of reacting the second intermediate by acyl transfer to cleave the acyl bond comprises hydrolyzing the acyl bond.  
     
     
         26 . The method of  claim 20 , wherein the substrate comprises an ester.  
     
     
         27 . The method of  claim 20 , wherein the substrate comprises an amide.