If you have an ACS member number, please enter it here so we can link this account to your membership. (optional)

ACS values your privacy. By submitting your information, you are gaining access to C&EN and subscribing to our weekly newsletter. We use the information you provide to make your reading experience better, and we will never sell your data to third party members.


Analytical Chemistry

New Tool For Phosphoproteomics

Antibody detects phosphorylated histidine independent of surrounding sequence

by Celia Henry Arnaud
June 3, 2013 | A version of this story appeared in Volume 91, Issue 22

Protein phosphorylation controls many cellular processes. Although four amino acids—serine, threonine, tyrosine, and histidine—can be targets for phosphorylation, good reagents for routine detection have been available for only three of the four. The lack of suitable reagents for histidine phosphorylation can be blamed on the chemical instability of the modification and the failure to raise good antibodies against it. Tom W. Muir and coworkers at Princeton University have used a stable phosphohistidine analog, phosphoryl-triazolylethylamine, to produce antibodies that detect in vitro and in vivo histidine phosphorylation independent of the surrounding protein sequence (Nat. Chem. Biol. 2013, DOI: 10.1038/nchembio.1259). They successfully used the antibody in various biological assays, including enzyme-linked immunosorbent assays (ELISAs), western blots, and immunoprecipitation. A survey of histidine phosphorylation in Escherichia coli cell lysates, which was done in conjunction with mass spectrometric analysis, revealed that the amount of phosphorylation depends on the carbon source and the availability of nitrogen in the growth medium. One drawback of the new antibody is minor cross-reactivity with phosphorylated tyrosine.


This article has been sent to the following recipient:

Chemistry matters. Join us to get the news you need.