Cell-based determination of HDAC10-mediated polyamine deacetylase activity.

Among histone deacetylases, HDAC10 is unique in its substrate preference for a specific acetylated polyamine, N-acetylspermidine (N-AcSpd), over other acetylated polyamines and peptides. As a polyamine deacetylase, HDAC10 catalyzes the conversion of N-AcSpd into spermidine, thereby enabling the cell to utilize this acetylated derivative to support polyamine homeostasis. Therefore, the level of HDAC10-mediated PDAC activity in a particular tissue and its exposure to extracellular N-AcSpd, a byproduct of certain intestinal microbes, may directly contribute to the maintenance of intracellular polyamine concentrations. This chapter provides detailed methods for determining relative levels of HDAC10-mediated polyamine deacetylase activity using cell-based assays. These cost-efficient methods are useful for identifying tissue-specific differences in PDAC activity and may also be adapted to enable high-throughput screening of effectors of HDAC10 function, such as HDAC inhibitors.