Oxidative stress increases ubiquitin--protein conjugates in synaptosomes.
Synaptosomes were incubated in the presence of FeSO4 to test the hypothesis that iron-catalyzed oxidative damage causes an increase in the ubiquitination of synaptosomal proteins. Incubation with 10 or 50 microM FeSO4 caused concentration-dependent increases in carbonyl groups (an indication of protein oxidation) and ubiquitinated proteins (determined by probing Western blots with a monoclonal antibody to ubiquitin). Differences in protein ubiquitination occurred within 5 min of incubation, indicating a rapid response to oxidative stress. Results of experiments with MG-132, an inhibitor of the degradation of ubiquitinated proteins, suggested that oxidative damage stimulated ubiquitination rather than inhibited degradation of ubiquitinated proteins. The data are consistent with the hypothesis that synaptic terminals utilize the ubiquitin/proteasome proteolytic pathway to degrade oxidatively damaged proteins.
Ramanathan
M
M
Department of Pharmacology, New Jersey Medical School UMDNJ, Newark 07103-2757, USA.
Hassanain
M
M
Levitt
M
M
Seth
A
A
Tolman
J S
JS
Fried
V A
VA
Ingoglia
N A
NA
D000818:Animals; D005296:Ferrous Compounds / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors; D014452:Ubiquitins / Q000378:metabolism*
D005296:Ferrous Compounds; D007976:Leupeptins; D009419:Nerve Tissue Proteins; D014452:Ubiquitins; C020748:ferrous sulfate; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde