TY - JOUR
T1 - Functional analysis of the spindle-checkpoint proteins using an in vitro ubiquitination assay.
AU - Tang, Zhanyun
AU - Yu, Hongtao
PY - 2004
Y1 - 2004
N2 - The spindle checkpoint helps to ensure the fidelity of chromosome segregation during mitosis and meiosis. In response to sister chromatids not properly attached to the mitotic spindle, this checkpoint blocks the activity of a large ubiquitin protein ligase complex, called the anaphase-promoting complex (APC) or cyclosome. This chapter describes the detailed protocols of an in vitro ubiquitination assay reconstituted with purified APC, cofactors, other ubiquitination enzymes, and the spindle-checkpoint proteins. This assay is extremely useful in dissecting the biochemical functions of various spindle-checkpoint proteins.
AB - The spindle checkpoint helps to ensure the fidelity of chromosome segregation during mitosis and meiosis. In response to sister chromatids not properly attached to the mitotic spindle, this checkpoint blocks the activity of a large ubiquitin protein ligase complex, called the anaphase-promoting complex (APC) or cyclosome. This chapter describes the detailed protocols of an in vitro ubiquitination assay reconstituted with purified APC, cofactors, other ubiquitination enzymes, and the spindle-checkpoint proteins. This assay is extremely useful in dissecting the biochemical functions of various spindle-checkpoint proteins.
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U2 - 10.1385/1-59259-811-0:227
DO - 10.1385/1-59259-811-0:227
M3 - Article
C2 - 15220533
AN - SCOPUS:4444336508
SN - 1064-3745
VL - 281
SP - 227
EP - 242
JO - Methods in molecular biology (Clifton, N.J.)
JF - Methods in molecular biology (Clifton, N.J.)
ER -