TY - CHAP
T1 - Studying protein ubiquitylation in yeast
AU - Hovsepian, Junie
AU - Becuwe, Michel
AU - Kleifeld, Oded
AU - Glickman, Michael
AU - Léon, Sébastien
N1 - Publisher Copyright: © Springer Science+Business Media New York 2016.
PY - 2016/9/1
Y1 - 2016/9/1
N2 - Ubiquitylation is a reversible posttranslational modifi cation that is critical for most, if not all, cellular processes and essential for viability. Ubiquitin conjugates to substrate proteins either as a single moiety (monoubiquitylation) or as polymers composed of ubiquitin molecules linked to each other with various topologies and structures (polyubiquitylation). This contributes to an elaborate ubiquitin code that is decrypted by specifi c ubiquitin-binding proteins. Indeed, these different types of ubiquitylation have different functional outcomes, notably affecting the stability of the substrate, its interactions, its activity, or its subcellular localization. In this chapter, we describe protocols to determine whether a protein is ubiquitylated, to identify the site that is ubiquitylated, and provide direction to study the topology of the ubiquitin modifi cation, in the yeast Saccharomyces cerevisiae.
AB - Ubiquitylation is a reversible posttranslational modifi cation that is critical for most, if not all, cellular processes and essential for viability. Ubiquitin conjugates to substrate proteins either as a single moiety (monoubiquitylation) or as polymers composed of ubiquitin molecules linked to each other with various topologies and structures (polyubiquitylation). This contributes to an elaborate ubiquitin code that is decrypted by specifi c ubiquitin-binding proteins. Indeed, these different types of ubiquitylation have different functional outcomes, notably affecting the stability of the substrate, its interactions, its activity, or its subcellular localization. In this chapter, we describe protocols to determine whether a protein is ubiquitylated, to identify the site that is ubiquitylated, and provide direction to study the topology of the ubiquitin modifi cation, in the yeast Saccharomyces cerevisiae.
KW - Histidine-tagged ubiquitin purifi cation
KW - Immunoprecipitation in denaturing conditions
KW - Ubiquitin
KW - Ubiquitin chain topology
KW - Ubiquitylation site mapping
KW - Yeast
UR - https://www.scopus.com/pages/publications/84987837804
U2 - 10.1007/978-1-4939-3756-1_5
DO - 10.1007/978-1-4939-3756-1_5
M3 - Chapter
C2 - 27613031
T3 - Methods in Molecular Biology
SP - 117
EP - 142
BT - Methods in Molecular Biology
ER -