Dual response of calpain to rat brain postdecapitative ischemia.
Calpains, Ca(2+)-dependent neutral proteinases (microM and mM Ca(2+)-sensitive), and their endogenous inhibitor calpastatin were examined in rat brain. Specific activity of m-calpain exceeded almost 10 times that of mu-calpain, and the both isoforms of calpain together with calpastatin were mainly located in the soluble fraction of homogenate. Acute postdecapitative ischemia of 15 min duration resulted in a gradual, time-dependent decrease of total mu-calpain activity (to 60% of control values) and in the moderate elevation of calpastatin activity (by 28%). The decrease of total mu-calpain activity coincided with its remarkable increase (above 300% of control values) in particulate fraction. In the case of m-calpain, the only observed effect of ischemia was its redistribution and, as a consequence, the elevation of activity in particulate fraction. The accumulation of breakdown products, resulting from calpain-catalyzed proteolysis of fodrin (as revealed by Western blotting) indicated activation of calpain under ischemia. The findings suggest that this rapid activation involves partial enzyme translocation toward membranes, and is followed (at least in acute phase) by mu-calpain downregulation and increased calpastatin activity.
Zalewska
T
T
Department of Neurochemistry, Medical Research Centre, Warsaw, Poland. bimol@ibbrain.ibb.war.pl
Zabłocka
B
B
Saido
T C
TC
Zajac
H
H
Domańska-Janik
K
K
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology*; D002545:Brain Ischemia / Q000201:enzymology*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002154:Calpain / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013049:Spectrin
D002135:Calcium-Binding Proteins; D002154:Calpain; D002352:Carrier Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D013049:Spectrin; C033668:calpastatin