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. 2010 May 21;3:12. doi: 10.3389/fnmol.2010.00012

Figure 4.

Figure 4

(A) Purity of isolated 26S proteasomes. Cytosolic (Cyt) and synaptic (Syn) 26S proteasomes were isolated using GST-UBL/His-UIM. In control (Con) experiments, GST was added instead of GST-UBL. Samples were resolved by 3–8% gradient native gel and silver-stained. (B) The same samples in (A) analyzed by SDS-PAGE, followed by silver staining. The control experiments showed only a few non-specific protein bands. (C) Activity of isolated proteasomes. Chymotrypsin-like peptidase activity of isolated 26S proteasomes resolved by 3–8% native gel was assayed with Suc-LLVY-AMC. (D) Differences in capped proteasome ratios. Silver-stained 26S proteasome bands in (C) were quantified by densitometry. The ratio of doubly- over singly-capped 26S was significantly higher in the cytosol (Cyt) than the synaptosome (Syn) (mean ± SEM, n = 4, *p < 0.05 by paired t-test).