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Deciphering Tissue-specific Ubiquitination by Mass Spectrometry

Protein ubiquitination is a highly conserved, central mechanism to regulate cellular events in all eukaryotes, such as proteasomal degradation, protein trafficking, DNA repair, synaptic plasticity and immune response. The consequence of protein ubiquitination is modulated by the structure of ubiquit...

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Formato: Artigo
Idioma:en
Publicado em: 2012
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Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC3475722/
https://ncbi.nlm.nih.gov/pubmed/22350876
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1007/978-1-61779-474-2_3
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Resumo:Protein ubiquitination is a highly conserved, central mechanism to regulate cellular events in all eukaryotes, such as proteasomal degradation, protein trafficking, DNA repair, synaptic plasticity and immune response. The consequence of protein ubiquitination is modulated by the structure of ubiquitin moiety attached on the substrates, including ubiquitin monomer and diverse polyubiquitin chains with different linkages (N-terminus, K6, K11, K27, K29, K33, K48 and K63). The development of ubiquitin-enrichment strategies coupled with sensitive mass spectrometry enables direct analysis of ubiquitinated proteins in cells, providing an invaluable tool for ubiquitin research. In this chapter we describe recent technology updates for analyzing tissue-specific ubiquitin conjugates in transgenic models, as well as targeted proteomics methods for quantifying different polyubiquitin chain linkages in any type of samples, including human tissues.