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

Expert Review of Proteomics
Yuanhui Ma, John R Yates

Abstract

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 ins...Continue Reading

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Citations

Jun 28, 2020·Nature Communications·Fleur KleinpenningAlbert J R Heck
Aug 11, 2020·Expert Review of Proteomics·Xingyun WuKui Wang
Sep 25, 2019·Nature Reviews. Microbiology·Christopher E LawsonKatherine D McMahon
Dec 11, 2020·Experimental Cell Research·Madison T Wright, Lars Plate
Jan 25, 2021·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Zhanwu HouHuadong Liu
May 1, 2020·Journal of Medicinal Chemistry·Emily J HananSteven T Staben
Sep 2, 2020·Journal of the American Chemical Society·Yanping ZhuDavid J Vocadlo
Aug 9, 2021·Current Opinion in Chemical Biology·Wouter van BergenMarc P Baggelaar

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