University of North Dakota

Masaru Miyagi

Chandra Sekhar Rao Kadiyala

K C Sekhar

Optimized enzymatic conditions incorporate a single oxygen atom into digested peptides using a peptidase. The incorporation of a single oxygen atom is especially useful for proteolyticO labeling in comparative proteomics. The optimized proteolyticO labeling minimizes the generation of a mixture of isotopic isoforms of the peptides resulting from incorporation of either one or twoO atoms. The outcome is accurate quantification of isotopically labeled peptides.

Method for single oxygen atom incorporation into peptides

A method for comparative proteomics using a peptidase under enzymatic conditions that permits the optimal incorporation of two oxygen atoms into a digested peptide. The method employs a peptidase to incorporate twoO atoms into a peptide set derived from a population of proteins at a conditioned state, which is compared to a second peptide set incorporated with a singleO atom derived from a population of proteins at a second conditioned state. Upon combining the two peptide sets, the populations of proteins are analyzed for qualitative and quantitative differences based on the content ofO atoms andO atoms in the digested peptides using mass spectrometry instrumentation. The method is advantageous to improve the efficiency and timeframe of peptidase catalyzedO labeling reactions which increased the accuracy and reliability of quantitative proteomic experiments.

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Grand Forks, ND, United States of America

Masaru Miyagi