Proteomics and pulse azidohomoalanine labeling of newly synthesized proteins: what are the potential applications?

Expert Rev Proteomics. 2018 Jul;15(7):545-554. doi: 10.1080/14789450.2018.1500902. Epub 2018 Jul 23.


Measuring the immediate changes in cells that arise from changing environmental conditions is crucial to understanding the underlying mechanisms involved. These changes can be measured with metabolic stable isotope fully labeled proteomes, but requires looking for changes in the midst of a large background. In addition, labeling efficiency can be an issue in primary and fully differentiated cells. Area covered: Azidohomoalanine (AHA), an analog of methionine, can be accepted by cellular translational machinery and incorporated into newly synthesized proteins (NSPs). AHA-NSPs can be coupled to biotin via CuAAC-mediated click-chemistry and enriched using avidin-based affinity purification. Thus, AHA-containing proteins or peptides can be enriched and efficiently separated from the whole proteome. In this review, we describe the development of mass spectrometry (MS) based AHA strategies and discuss their potential to measure proteins involved in immune response, secretome, gut microbiome, and proteostasis as well as their potential for clinical uses. Expert commentary: AHA strategies have been used to identify synthesis activity and to compare two biological conditions in various biological model organisms. In combination with instrument development, improved sample preparation and fractionation strategies, MS-based AHA strategies have the potential for broad application, and the methods should translate into clinical use.

Keywords: Azidohomoalanine; Biological studies; Proteomics; application; newly synthesized protein.

Publication types

  • Research Support, N.I.H., Extramural
  • Review

MeSH terms

  • Alanine / analogs & derivatives*
  • Alanine / metabolism
  • Animals
  • Humans
  • Immunity
  • Isotope Labeling*
  • Microbiota
  • Protein Biosynthesis*
  • Proteomics / methods*
  • Proteostasis


  • azidohomoalanine
  • Alanine