Journals of Gerontology Series A: Biological Sciences and Medical Sciences Large Type Edition
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The Journals of Gerontology Series A: Biological Sciences and Medical Sciences 55:B220-B227 (2000)
© 2000 The Gerontological Society of America

Increase of Oxidatively Modified Protein Is Associated With a Decrease of Proteasome Activity and Content in Aging Epidermal Cells

Isabelle Petropoulosa, Mariangela Conconia, Xin Wangb, Batya Hoenelb, François Brégégèrea,b, Yoram Milnerb and Bertrand Frigueta

a Laboratoire de Biologie et Biochimie Cellulaire du Vieillissement, Université Denis Diderot, Paris, France
b Department of Biological Chemistry, The Hebrew University, Jerusalem, Israel

Correspondence: Bertrand Friguet, Laboratoire de Biologie et Biochimie Cellulaire du Vieillissement, Universit\|[eacute]\| Denis Diderot-Paris 7, CC 7128, 2 place Jussieu, 75251 Paris Cedex 05, France E-mail: bfriguet{at}paris7.jussieu.fr.

Decision Editor: Jay Roberts, PhD

For the process of aging in epidermal cells to be characterized, the status of oxidized and damaged protein accumulation and removal by the proteasome has been investigated. Modified protein content and proteasome activity were assayed in lysates of epidermal cells from donors of different ages. Increased levels of oxidized proteins, glycated proteins, and proteins modified by the lipid peroxidation product 4-hydroxy-2-nonenal were observed in cells from old donors. At the same time, a decline of chymotrypsin-like and peptidylglutamyl-peptide hydrolase activities of the proteasome was found in aging keratinocytes. This age-related decline of the proteasome peptidase activities can be explained, at least in part, by a decreased proteasome content as observed by immunoblotting and enzyme-linked immunosorbent assay. In keratinocyte cultures, a decrease of proteasome activity and content was observed upon serial passaging. In cultures, as well as in skin, an inverse relationship was found between the aging marker ß-galactosidase and the proteasome content. These results suggest that proteasome is downregulated during replicative senescence as well as in aged cells in vivo, possibly resulting in the accumulation of modified proteins.




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