The Immunoproteasome in Human Lens Epithelial Cells During Oxidative Stress.
OBJECTIVE
The immunoproteasome is known to generate peptides for antigen presentation. However, it has also been proposed to have additional functions such as stress response. The propensity of the immunoproteasome for degradation of oxidatively damaged proteins and peptides makes it interesting in the context of cataract formation and prevention. This study hypothesized that the immunoproteasome is present in human cataractous lenses and that oxidative stress will induce its expression, affect its proteolytic activity and its intracellular location in native cultured human lens epithelial cells (HLECs).


METHODS
The expression of the immunoproteasome and the constitutive proteasome subunits β1i/β1, β2i/β2, and β5i/β5 were studied by using Western blotting. The chymotrypsin-like activity was investigated for possible oxidative stress response. Inhibitors specific for the immuno- and constitutive proteasome, ONX-0914 and MG-132, respectively, were used to study their relative contributions to total proteasome activity. The intracellular location of the proteasomal subunits β5i and β5 was studied by immunocytochemistry.


RESULTS
Immunoproteasome subunits were detected both in the lens epithelium and in the lens fibers derived from cataract surgery. Oxidative stress to cultured HLECs upregulated the immunoproteasome but not the constitutive proteasome. The chymotrypsin-like activity decreased with increased oxidative stress and the two proteasome types contributed equally to total proteasome activity. Immunocytochemical labeling of subunit β5i showed mainly cytosolic localization, whereas subunit β5 was localized predominantly in the nucleus. H2O2-induced challenge increased the expression of the immunoproteasome.


CONCLUSIONS
The present findings indicate a role for the immunoproteasome in oxidative stress management in the lens.
Department of Clinical Neuroscience, Institute of Neuroscience and Physiology, the Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
Anne
A
Petersen
Department of Clinical Neuroscience, Institute of Neuroscience and Physiology, the Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden.
Madeleine
M
Zetterberg