Purification of the mitotic checkpoint complex, an inhibitor of the APC/C from HeLa cells

Methods in Molecular Biology
Valery Sudakin, Tim J Yen

Abstract

The mitotic checkpoint prevents cells with unaligned chromosomes from prematurely exiting mitosis by inhibiting the anaphase-promoting complex/cyclosome (APC/C) from targeting key proteins for ubiquitin-mediated proteolysis. Great efforts have been made recently to identify and characterize the factors involved in this checkpoint pathway. These studies will ultimately lead to a better understanding of cell cycle defects and chromosome instability. We report here a protocol for purification of an inhibitor of the APC/C from HeLa cells, called mitotic checkpoint complex (MCC). Our procedure is based on biochemical purification and characterization of the APC/C inhibitory activity from extracts of HeLa cells.

Citations


❮ Previous
Next ❯

Related Concepts

Related Feeds

Cell Checkpoints & Regulators

Cell cycle checkpoints are a series of complex checkpoint mechanisms that detect DNA abnormalities and ensure that DNA replication and repair are complete before cell division. They are primarily regulated by cyclins, cyclin-dependent kinases, and the anaphase-promoting complex/cyclosome. Here is the latest research.