Structural changes in 20S proteasomes after HAMLET treatment <i>in vitro</i>.

Abstract

<p>(A, B) <i>In vitro</i> fragmentation of human 20S proteasomes after 20 minutes incubation with HAMLET (7 µM), compared to the α-lactalbumin control. (A) The proteins were separated by SDS-PAGE and gels were stained with Amido black. Changes in 20S proteasome subunits are indicated by arrows. The subunits identified by mass spectrometry are shown in the table. (B) Western Blot of samples from <i>in vitro</i> mixing experiment with 20S proteasomes and HAMLET or α-lactalbumin using antibodies against the 20S proteasome core subunits. (C) MALDI-TOF mass spectrometry of intact 20S proteasomes (top panel) and major changes after HAMLET treatment (bottom panel), indicated by arrows.</p