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. 2023 May 11;299(6):104811. doi: 10.1016/j.jbc.2023.104811

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© 2023 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

Blinded brain samples from a cohort of human healthy controls and AD patients exhibit 20S proteasome activity that is positively correlated with the severity of the AD state.A, purified human 20/26S proteasomes or tissue lysates from human parahippocampal gyrus (PHG) of unaffected (C), preclinical AD-diagnosed (P-AD), and AD-diagnosed individuals were incubated with MV151. Samples were treated under non-denaturing conditions and run on 4% native gels, which were imaged with the Cy3 channel. B, imaged gels were then immunoblotted with antibodies to the indicated core proteasome subunit (β5). Cy3 and immunoblot bands correspond to individual proteasome complexes (30S, 26S, 20S; asterisks for purified proteasomes and arrows for tissue homogenate). Note: we predominantly detect 20S proteasomes. C, multiple variable correlation analysis with Tau band intensity normalized to actin, 20S proteasome MV151 Cy3 signal normalized to β5 band intensity (activity), and Braak Stage as variables in PHG is shown. The significance table shows the two-tailed p-value with a confidence interval of 95% as a correlation matrix. D, multiple variable correlation analysis with Tau band intensity normalized to actin, 20S proteasome β5 band intensity (abundance), and Braak Stage as variables in PHG is shown. The significance table shows the two-tailed p-value with a confidence interval of 95% as a correlation matrix. E, Cy3 in-gel visualization of MV151 of purified human 20S/26S proteasomes or tissue lysates from human visual cortex (VC) of unaffected (C), preclinical AD-diagnosed (P-AD), and AD-diagnosed individuals were treated as in (A). F, imaged gels were then immunoblotted with antibodies to the indicated core proteasome subunit (β5). Cy3 and immunoblot bands correspond to individual proteasome complexes (30S, 26S, 20S) (arrows). Note: we predominantly detect 20S proteasomes. G, multiple variable correlation analysis with Tau band intensity normalized to actin, 20S proteasome MV151 Cy3 signal normalized to β5 band intensity (activity), and Braak Stage as variables in VC is shown. Statistics of the correlation analysis are shown below the graph. H, multiple variable correlation analysis with Tau band intensity normalized to actin, 20S proteasome β5 band intensity (abundance), and Braak Stage as variables in VC is shown with statistics shown below the graph. See also Fig. S6.