A substrate for proteasomes
Ac-Nle-Pro-Nle-Asp-AMC is a substrate for proteasomes that is cleaved by the caspase-like site, also known as postglutamyl peptide hydrolase (PGPH), with specific activities of 113 and 6.6 nmol/min/mg by the 26S (rabbit muscle) and 20S (yeast) proteasomes, respectively. Caspase-like activity can be quantified by fluorescent detection of free AMC (also known as 7-amino-4-methylcoumarin), which is excited at 340-360 nm and emits at 440-460 nm. Ac-Nle-Pro-Nle-Asp-AMC inhibits the chymotrypsin-like activity of the proteasome allosterically when bound to the caspase-like site. It has been used to determine L-DOPA’s effects on protein turnover rates.
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