RT Journal Article
T1 Proteomic Approaches to Identifying Carbonylated Proteins in Brain Tissue
A1 Linares Gómez, María
A1 Marín-García, Patricia
A1 Méndez, Darío
A1 Puyet Catalina, Antonio
A1 Díez Martín, Amalia
A1 Bautista Santa Cruz, José Manuel
AB Oxidative stress plays a critical role in the pathogenesis of a number of diseases. The carbonyl end products of protein oxidation are among the most commonly measured markers of oxidation in biological samples. Protein carbonyl functional groups may be derivatized with 2,4-dinitrophenylhydrazine (DNPH) to render a stable 2,4-dinitrophenylhydrazone-protein (DNP-protein) and the carbonyl contents of individual proteins then determined by two-dimensional electrophoresis followed by immunoblotting using specific anti-DNP antibodies. Unfortunately, derivatization of proteins with DNPH could affect their mass spectrometry (MS) identification. This problem can be overcome using nontreated samples for protein identification. Nevertheless, derivatization could also affect their mobility, which might be solved by performing the derivatization step after the initial electrophoresis. Here, we compare two-dimensional redox proteome maps of mouse cerebellum acquired by performing the DNPH derivatization step before or after electrophoresis and detect differences in protein patterns. When the same approach is used for protein detection and identification, both methods were found to be useful to identify carbonylated proteins. However, whereas pre-DNPH derivatized proteins were successfully analyzed, high background staining complicated the analysis when the DNPH reaction was performed after transblotting. Comparative data on protein identification using both methods are provided.
PB American Chemical Society
SN 1535-3893
SN 1535-3907
YR 2011
FD 2011
LK https://hdl.handle.net/20.500.14352/94598
UL https://hdl.handle.net/20.500.14352/94598
LA eng
NO J. Proteome Res. 2011, 10, 4, 1719–1727Publication Date:January 15, 2011 https://doi.org/10.1021/pr101014e
NO Ministerio de Educación (España)
NO Ministerio de Ciencia e Innovación (España)
NO Comunidad de Madrid
DS Docta Complutense
RD 11 abr 2025