RT info:eu-repo/semantics/bookPart
T1 In vivo quantitative proteomics for the study of oncometabolism.
A1 Hernández Fernaud, Juan Ramón
A1 Reid, Steven
A1 Zanivan, Sara
A2 BioquímicaMicrobiología, Biología Celular y Genética
A2 The Beatson Institute for Cancer ResearchGlasgow, UK
AB The active reprograming of cellular metabolism is a primary driver of oncogenesis and ahallmark of established neoplastic lesions. Much of this reprogramming depends on theexpression levels and posttranslational modifications (PTMs) of metabolic enzymes.Stable isotope labeling of amino acids in culture (SILAC) is an amino acid-based labelingtechnique that can be used both in vitro and in vivo to comparatively assess the levelsand PTMs of proteins. To this aim, SILAC-labeled cell lysates can be spiked into eachsample as a standard, followed by the analysis of specimens by mass spectrometry(MS). Combined with appropriate protocols for the lysis and preparation of samplesfor MS, this technique allows for the accurate and in-depth quantification of the proteomeof a wide variety of cell and tissue samples. In particular, SILAC can be employed toinfer the metabolic state of neoplastic lesions and obtain a profound understanding ofthe proteomic alterations that accompany oncogenesis and tumor progression. Here, we describe a proteomic approach based on SILAC, high-resolution chromatographyand high-accuracy MS for comparing levels and phosphorylation status of proteinsbetween the samples of interest. This method can be applied not only to the proteomicstudy of oncometabolism in murine tissues, but also to the study of cellular samples andhuman specimens.
SN 978-0-12-801329-8
YR 2014
FD 2014
LK http://riull.ull.es/xmlui/handle/915/38822
UL http://riull.ull.es/xmlui/handle/915/38822
LA en
DS Repositorio institucional de la Universidad de La Laguna
RD 19-oct-2024