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Liquid- And Gas-Phase Nitration of Bovine Serum Albumin Studied by LC-MS and LC-MS/MS Using Monolithic Columns

Liquid- And Gas-Phase Nitration of Bovine Serum Albumin Studied by LC-MS and LC-MS/MS Using Monolithic Columns

Wolfgang Walcher, Thomas Franze, Michael G Weller, Ulrich Pöschl, Christian G Huber

J Proteome Res. Sep-Oct 2003;2(5):534-42.

PMID: 14582650

Abstract:

Post-translational nitration of proteins was analyzed by capillary reversed-phase high-performance liquid chromatography (RP-HPLC) on-line interfaced to electrospray ionization mass spectrometry (ESI--MS) or tandem mass spectrometry (ESI--MS/MS). Both methods were compared using a tryptic digest of bovine serum albumin (BSA) and yielded sequence coverages of 95% and 33% with RP-HPLC--ESI--MS and RP-HPLC--ESI--MS/MS, respectively. At least 95% of the tyrosines were covered by the former method, whereas the latter method only detected less than 50% of the tyrosine-containing peptides. Upon liquid-phase nitration of BSA in aqueous solution using an excess of tetranitromethane, at least 16 of the 20 tyrosine residues were found to be nitrated. After exposure of solid BSA samples to gaseous nitrogen dioxide and ozone at atmospherically relevant concentration levels, only 3 nitrated peptides were detected. By use of such a model system, RP-HPLC--ESI--MS proved to be a rapid and highly efficient method for the comprehensive and quantitative detection of protein nitration.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42411674	Albumin, Nitrated from bovine serum			Price

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