Method for identifying activation of transferases
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United States of America Patent
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Aug 29, 2017
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Oct 18, 2011
filing date -
Oct 20, 2010
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Abstract
The present invention provides a method for identifying differential activation of a bisubstrate protein modifying enzyme between samples, comprising:
- (i) incubating a first sample with x different concentrations of the non-protein substrate of said enzyme, wherein x is 2 or greater than 2;(ii) quantifying modification of a polypeptide in said sample at each of the x different concentrations of the non-protein substrate;(iii) determining the affinity of said enzyme for said non-protein substrate;(iv) repeating steps (i) to (iii) for a second or subsequent sample; and(v) comparing the affinity of said enzyme for said non-protein substrate between said samples;wherein a difference in affinity of said enzyme for said non-protein substrate between samples is indicative of differential activation of said enzyme between samples. The present invention also provides a method for identifying an in vivo substrate of a bisubstrate protein modifying enzyme.
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