Method and Apparatus for Measuring Protein Post-Translational Modification

Abstract

The present invention includes a method for analyzing reactions. The method includes the steps of providing a solution of at least one acceptor chemical and at least one donor chemical. The donor chemical is capable of donating a chemical moiety to the acceptor chemical. The solution further includes at least one controller chemical that affects the reaction between the donor chemical and the acceptor chemical. The solution is then incubated so that a portion of the acceptor chemical reacts with the donor chemical to form an acceptor product. Unreacted donor chemical is separated from the acceptor product. The acceptor product or the donor chemical is then measured using X-ray fluorescence. Another aspect of the present invention includes a method for analyzing protein function. The method includes the steps of providing a solution of at least one acceptor chemical and at least one donor chemical. The donor chemical is capable of donating a chemical moiety to the acceptor chemical. The donor chemical includes a functional group selected from ester, anhydride, imide, acyl halide, and amide. The solution is then incubated so that a portion of the acceptor chemical reacts with the donor chemical to form an acceptor product. Unreacted donor chemical is separated from the acceptor product. The acceptor product or the donor chemical is then measured using X-ray fluorescence. Yet another aspect of the present invention includes a method for analyzing protein function. The method includes the steps of providing a solution of at least one acceptor chemical and at least one donor chemical. The solution is then incubated so that a portion of the acceptor chemical reacts with the donor chemical to form an acceptor product. Unreacted donor chemical is separated from the acceptor product. The acceptor product or the donor chemical is then measured using X-ray fluorescence. An additional analytical method is also used to measure either the acceptor product or the donor chemical.

1 . A method for analyzing reactions, comprising:
 a. Providing a solution comprising at least one acceptor chemical and at least one donor chemical, wherein the donor chemical is capable of donating a chemical moiety comprising a chemical element to the acceptor chemical, and at least one controller chemical capable of affecting the reaction between the donor chemical and the acceptor chemical;   b. Incubating the solution to allow a portion of the acceptor chemical to react with the donor chemical to form an acceptor product;   c. Separating at least a portion of the unreacted donor chemical from the acceptor product; and   d. Measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical.   
     
     
         2 . The method of  claim 1 , wherein the controller chemical comprises at least one of the species selected from the list protein, amino acid, peptide, catalyst; carbohydrate, metal, lipid, nucleotide, polynucleotide, reactive oxygen species, and reactive nitrogen species. 
     
     
         3 . The method of  claim 2 , wherein the donor chemical comprises a chemical element having an atomic number greater than eight. 
     
     
         4 . The method of  claim 3 , wherein the solution further comprises at least one additional controller chemical. 
     
     
         5 . The method of  claim 4 , wherein the acceptor molecule comprises a chemical selected from the list water; a chemical comprising at least one functional group selected from the list peptide, amino acid, alcohol, alkoxide, aryloxide, aromatic alcohol, amine, thiol, thiolate, nucleotide; and combinations thereof. 
     
     
         6 . The method of  claim 5 , wherein the donor chemical comprises a functional group selected from the list of anhydride, ester, amide, imide, acyl halide, and combinations thereof. 
     
     
         7 . The method of  claim 6 , further comprising the step of removing at least a portion of the solvent before measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical. 
     
     
         8 . The method of  claim 7 , wherein the acceptor chemical comprises at least one chemical element having an atomic number greater than eight and which is not donated from the donor chemical to the acceptor chemical. 
     
     
         9 . The method of  claim 8 , wherein the solution comprises at least one chemical selected from the list of donor chemical and acceptor chemical is present in a concentration less than about 100 millimolar. 
     
     
         10 . A method for analyzing protein function, comprising:
 a. Providing a solution comprising at least one acceptor chemical and at least one donor chemical, where the donor chemical is capable of donating a chemical moiety comprising a chemical element to the acceptor chemical, wherein the donor chemical comprises a functional group selected from ester, anhydride, imide, acyl halide, amide, and combinations thereof;   b. Incubating the solution to allow a portion of the acceptor chemical to react with the donor chemical to form an acceptor product;   c. Separating at least a portion of the unreacted donor chemical from the acceptor product; and   d. Measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical.   
     
     
         11 . The method of  claim 10 , further comprising at least one controller chemical capable of affecting the reaction between the donor chemical and the acceptor chemical. 
     
     
         12 . The method of  claim 11 , wherein a covalent bond is formed during the reaction of the donor chemical and the acceptor chemical. 
     
     
         13 . The method of  claim 12 , wherein the solution further comprises at least one additional controller chemical. 
     
     
         14 . The method of  claim 13 , wherein the donor chemical comprises a chemical element having an atomic number greater than eight. 
     
     
         15 . The method of  claim 14 , further comprising the step of removing at least a portion of the solvent before measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical. 
     
     
         16 . A method for analyzing protein function, comprising:
 a. Providing a solution comprising at least one acceptor chemical and at least one donor chemical, where the donor chemical is capable of donating a chemical moiety comprising a chemical element to the acceptor chemical;   b. Incubating the solution to allow a portion of the acceptor chemical to react with the donor chemical to form an acceptor product;   c. Separating at least a portion of the unreacted donor chemical from the product chemical;   d. Measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical; and   e. Measuring using at least one additional analytical method at least one of the chemicals selected from the list of acceptor product and the donor chemical.   
     
     
         17 . The method of  claim 16 , wherein the solution further comprises at least one controller chemical capable of affecting the reaction between the donor chemical and the acceptor chemical. 
     
     
         18 . The method of  claim 17 , wherein the solution further comprises at least one additional controller chemical. 
     
     
         19 . The method of  claim 18 , wherein the donor chemical comprises a chemical element having an atomic number greater than eight. 
     
     
         20 . The method of  claim 19 , further comprising the step of removing at least a portion of the solvent before measuring using x-ray fluorescence at least one of the chemicals selected from the list of acceptor product and the donor chemical.