Proteasome Active Sites Allosterically Regulate Each Other, Suggesting a Cyclical Bite-Chew Mechanism for Protein Breakdown》.pdf

Molecular Cell, Vol. 4, 395–402, September, 1999, Copyright 1999 by Cell Press Proteasome Active Sites Allosterically Regulate Each Other, Suggesting a Cyclical Bite-Chew Mechanism for Protein Breakdown Alexei F. Kisselev, Tatos N. Akopian,† (Groll et al., 1997; Baumeister et al., 1998). Each outer Vanesa Castillo,‡ Alfred L. Goldberg* ring contains seven different subunits, while the two Department of Cell Biology inner rings each contain seven different subunits. Harvard Medical School Three of these subunits harbor active sites that face Boston, Massachusetts 02115 the inner cavity of the cylinder (Groll et al., 1997; Dick et al., 1998). These active sites are classified based on their specificity toward model peptide substrates. Two Summary chymotrypsin-like sites cleave preferentially after large hydrophobic residues, two trypsin-like sites cut after In eukaryotes, the 20S proteasome contains two chy- basic residues, while two sites, which are usually termed motrypsin-like, two trypsin-like, and two active sites “peptidylglutamyl-peptide hydrolyzing” (PGPH) cleave shown here to have caspase-like specificity. We report after acidic residues (Chen and Hochstrasser, 1996; Ar- that certain sites allosterically regulate each other’s endt and Hochstrasser, 1997; Groll et al., 1997; Heine- activities. Substrates of a chymotrypsin-like site stim-