Proteasomal degradation of beta-carotene metabolite--modified proteins.
Free radical attack on beta-carotene results in the formation of high amounts of carotene breakdown products (CBPs) having biological activities. As several of the CBPs are reactive aldehydes, it has to be considered that these compounds are able to modify proteins. Therefore, the aim of the study was to investigate whether CBP-modification of proteins is leading to damaged proteins recognized and degraded by the proteasomal system. We used the model proteins tau and ferritin to test whether CBPs will modify them and whether such modifications lead to enhanced proteasomal degradation. To modify proteins, we used crude CBPs as a mixture obtained after hypochloric acid derived BC degradation, as well as several single compounds, as apo8'-carotenal, retinal, or beta-ionone. The majority of the CBPs found in our reaction mixture are well known metabolites as described earlier after BC degradation using different oxidants. CBPs are able to modify proteins, and in in vitro studies, we were able to demonstrate that the 20S proteasome is able to recognize and degrade CBP-modified proteins preferentially. In testing the proteolytic response of HT22 cells toward CBPs, we could demonstrate an enhanced protein turnover, which is sensitive to lactacystin. Interestingly, the proteasomal activity is resistant to treatment with CBP. On the other hand, we were able to demonstrate that supraphysiological levels of CBPs might lead to the formation of protein-CBP-adducts that are able to inhibit the proteasome. Therefore, the removal of CBP-modified proteins seems to be catalyzed by the proteasomal system and is effective, if the formation of CBPs is not overwhelming and leading to protein aggregates.
Department of Pediatric Pulmonology, Children's University Hospital III, Heidelberg, Germany.
Olaf
O
Sommerburg
Nicole
N
Karius
Werner
W
Siems
Claus-Dieter
CD
Langhans
Michael
M
Leichsenring
Nicolle
N
Breusing
Tilman
T
Grune
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D005293:Ferritins / Q000187:drug effects / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D051379:Mice; D018384:Oxidative Stress; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D019207:beta Carotene / Q000378:metabolism*; D016875:tau Proteins / Q000187:drug effects / Q000378:metabolism*
D061988:Proteasome Inhibitors; D016875:tau Proteins; D019207:beta Carotene; C067713:lactacystin; D005293:Ferritins; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine