Enzymatic properties of the proteasome purified from starfish oocytes and its catalytic subunits involved in oocyte maturation.
The 20S proteasome was purified from oocytes of the starfish Asterina pectinifera and its enzymatic properties were investigated. The chymotrypsin-like activities were potently inhibited by PSI as well as MG115, whereas the trypsin-like and peptidyl-glutamyl peptide-hydrolyzing (PGPH) activities were not or only weakly inhibited by PSI and MG115. The inhibitory ability of MG115 toward germinal vesicle breakdown (GVBD) coincided with those toward the trypsin-like and PGPH activities, and PSI showed no inhibitory effect on GVBD. We have previously reported that the inhibition pattern toward GVBD of peptidyl-argininals, which potently inhibited the proteasomal trypsin-like activity rather than the chymotrypsin-like activity, correlated with the inhibition pattern toward the chymotrypsin-like activity of the proteasome. These results, together with the peptidyl-argininals scarcely inhibiting the PGPH activity at concentrations sufficient for the inhibition toward GVBD, indicate that both the chymotrypsin-like and trypsin-like activities, but not the PGPH activity, of the proteasome are responsible for degradation of the physiological substrate during starfish oocyte maturation. It was also suggested that the inhibition of a single catalytic site of the proteasome is not sufficient for prevention of the proteasomal function.
Tanaka
E
E
Department of Biochemistry, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo 060-0812 , Japan.
Takagi Sawada
M
M
Sawada
H
H
D000818:Animals; D002384:Catalysis; D002453:Cell Cycle; D002454:Cell Differentiation / Q000502:physiology*; D002918:Chymotrypsin / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D005260:Female; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000494:pharmacology*; D009865:Oocytes / Q000737:chemistry* / Q000254:growth & development*; D046988:Proteasome Endopeptidase Complex; D013215:Starfish / Q000502:physiology*; D014357:Trypsin / Q000378:metabolism
D009097:Multienzyme Complexes; D002918:Chymotrypsin; D014357:Trypsin; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex