Changes in proteasome activity following transient ischemia.
Succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide (Suc-LLVY-MCA) hydrolyzing activities of the 20S and 26S proteasomes in the gerbil cortex following transient forebrain ischemia were examined. Using extraction solutions without ATP, only 20S proteasome activity was noted after separation with glycerol gradient centrifugation. When these extracts were incubated with ATP and an ATP-regenerating system prior to glycerol gradient separation, both 20S and 26S proteasome activities were detected. Following 10 min of ischemia, the activity of the 26S proteasomes decreased, whereas the 20S proteasome activity increased after 30 min of reperfusion. These changes returned to the control level after 1 h. The active 26S proteasomes were formed with ATP-dependent association with the 20S proteasomes and several subunits and the 26S proteasomes degraded ubiquitin-protein conjugates. These results indicate that proteasome activity might not be irreversibly impaired after transient ischemia. However, transient inhibition of ATP-dependent conversion of 20S to 26S proteasomes in vitro must be one of the causes of the accumulation of the ubiquitin-protein conjugates in the early reperfusion period.
Kamikubo
T
T
Department of Neurosurgery, Jikei University School of Medicine, Tokyo, Japan.
Hayashi
T
T
D000255:Adenosine Triphosphate / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D002499:Centrifugation, Density Gradient; D003374:Coumarins / Q000378:metabolism; D005260:Female; D005849:Gerbillinae; D015151:Immunoblotting; D002546:Ischemic Attack, Transient / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D015388:Organelles / Q000378:metabolism*; D015424:Reperfusion
D003374:Coumarins; D009842:Oligopeptides; D000255:Adenosine Triphosphate; C046582:succinyl-leucyl-leucyl-valyl-tyrosyl-methylcoumarinamide