Unknown

Dataset Information

0

A chemical approach for detecting sulfenic acid-modified proteins in living cells.


ABSTRACT: Oxidation of the thiol functional group in cysteine (Cys-SH) to sulfenic (Cys-SOH), sulfinic (Cys-SO2H) and sulfonic acids (Cys-SO3H) is emerging as an important post-translational modification that can activate or deactivate the function of many proteins. Changes in thiol oxidation state have been implicated in a wide variety of cellular processes and correlate with disease states but are difficult to monitor in a physiological setting because of a lack of experimental tools. Here, we describe a method that enables live cell labeling of sulfenic acid-modified proteins. For this approach, we have synthesized the probe DAz-1, which is chemically selective for sulfenic acids and cell permeable. In addition, DAz-1 contains an azide chemical handle that can be selectively detected with phosphine reagents via the Staudinger ligation for identification, enrichment and visualization of modified proteins. Through a combination of biochemical, mass spectrometry and immunoblot approaches we characterize the reactivity of DAz-1 and highlight its utility for detecting protein sulfenic acids directly in mammalian cells. This novel method to isolate and identify sulfenic acid-modified proteins should be of widespread utility for elucidating signaling pathways and regulatory mechanisms that involve oxidation of cysteine residues.

SUBMITTER: Reddie KG 

PROVIDER: S-EPMC3529510 | biostudies-literature | 2008 Jun

REPOSITORIES: biostudies-literature

altmetric image

Publications

A chemical approach for detecting sulfenic acid-modified proteins in living cells.

Reddie Khalilah G KG   Seo Young Ho YH   Muse Iii Wilson B WB   Leonard Stephen E SE   Carroll Kate S KS  

Molecular bioSystems 20080314 6


Oxidation of the thiol functional group in cysteine (Cys-SH) to sulfenic (Cys-SOH), sulfinic (Cys-SO2H) and sulfonic acids (Cys-SO3H) is emerging as an important post-translational modification that can activate or deactivate the function of many proteins. Changes in thiol oxidation state have been implicated in a wide variety of cellular processes and correlate with disease states but are difficult to monitor in a physiological setting because of a lack of experimental tools. Here, we describe  ...[more]

Similar Datasets

| S-EPMC171382 | biostudies-literature
| S-EPMC3587177 | biostudies-literature
| S-EPMC2526167 | biostudies-literature
| S-EPMC8321940 | biostudies-literature
| S-EPMC7751106 | biostudies-literature
| S-EPMC2741475 | biostudies-literature
| S-EPMC4064372 | biostudies-literature
| S-EPMC3946320 | biostudies-literature
| S-EPMC5775914 | biostudies-literature
| S-EPMC4184475 | biostudies-literature