Specific immunoadsorption of the high molecular weight multicatalytic protease, macropain, from postmicrosomal extracts of BHK fibroblasts inhibited ATP-dependent proteolysis of exogenous protein substrates. The immunoprecipitated macropain represented the latent (L) form of the protease because it had low protease activity but was activated by methods that activate purified macropain L. Reconstitution of the antibody-treated extracts with purified macropain L, but not macropain A, from bovine heart or human erythrocytes, completely restored ATP-dependent proteolysis, even though ATP did not directly activate either purified macropain L or the immunoprecipitated protease. Reconstituted ATP-dependent proteolysis was saturable with respect to added macropain and never exceeded the level of proteolysis present in the original extract. These results indicate that macropain L plays a key role in ATP-dependent proteolysis but suggest that the protease may require interaction with or modification by another cellular component to demonstrate this effect.
|Original language||English (US)|
|Number of pages||6|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - Apr 28 1989|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology