Unknown

Dataset Information

0

Biotinylated probes for the analysis of protein modification by electrophiles.


ABSTRACT: Formation of covalent protein adducts by lipid electrophiles contributes to diseases and toxicities linked to oxidative stress, but analysis of the adducts presents a challenging analytical problem. We describe selective adduct capture using biotin affinity probes to enrich protein and peptide adducts for analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). One approach employs biotinamidohexanoic acid hydrazide to covalently label residual carbonyl groups on adducts. The other employs alkynyl analogs of lipid electrophiles, which form adducts that can be postlabeled with azidobiotin tags by Cu(+)-catalyzed cycloaddition (Click chemistry). To enhance the selectivity of adduct capture, we use an azidobiotin reagent with a photocleavable linker, which allows recovery of adducted proteins and peptides under mild conditions. This approach allows both the identification of protein targets of lipid electrophiles and sequence mapping of the adducts.

SUBMITTER: Codreanu SG 

PROVIDER: S-EPMC3811082 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

altmetric image

Publications

Biotinylated probes for the analysis of protein modification by electrophiles.

Codreanu Simona G SG   Kim Hye-Young H HY   Porter Ned A NA   Liebler Daniel C DC  

Methods in molecular biology (Clifton, N.J.) 20120101


Formation of covalent protein adducts by lipid electrophiles contributes to diseases and toxicities linked to oxidative stress, but analysis of the adducts presents a challenging analytical problem. We describe selective adduct capture using biotin affinity probes to enrich protein and peptide adducts for analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS). One approach employs biotinamidohexanoic acid hydrazide to covalently label residual carbonyl groups on adducts. The other  ...[more]

Similar Datasets

| S-EPMC6174696 | biostudies-literature
| S-EPMC1149093 | biostudies-other
| S-EPMC2636689 | biostudies-literature
| S-EPMC2602764 | biostudies-literature
| S-EPMC3805131 | biostudies-literature
| S-EPMC5755618 | biostudies-literature
| S-EPMC3742165 | biostudies-literature
| S-EPMC2693349 | biostudies-literature
| S-EPMC3579456 | biostudies-literature
| S-EPMC3544294 | biostudies-literature