Identification of gene family of caspases in rat kidney and altered expression in ischemia-reperfusion injury.
In the present study, we demonstrate that rat kidney contains caspase activity that was markedly inhibited by specific peptide inhibitors of caspases but not by inhibitors of Ser, Cys, Asp, or metalloproteinases. Using primers based on the nucleotide sequence of known members of Ced-3/interleukin-1 beta-converting enzyme (ICE) family from human origin, we have identified by reverse-transcription (RT) polymerase chain reaction (PCR) analyses that rat kidney transcribes the genes for caspase-1 (ICE), caspase-2 (Nedd2), caspase-3 (CPP32), and caspase-6 (Mch2). RT-PCR products, when subcloned and sequenced, provided full-length cDNAs for ICE (1,209 bp) and CPP32 (786 bp) and partial cDNA products for Mch2 (561 bp) and Nedd2 (811 bp). The sequence analysis of the caspase cDNAs showed conserved catalytic site QACRG as well as Asp cleavage site. Rat kidneys subjected to ischemia-reperfusion injury revealed differential expression of caspases with marked increase in CPP32 and ICE mRNA and proteins during reperfusion, transient increase in Nedd2 mRNA and proteins during ischemia and the early period of reperfusion, and little change in Mch2 expression during the ischemia or reperfusion period. The altered expression suggests that caspases may act in concert in a cascade and may play an important role in ischemic acute renal failure.
Kaushal
G P
GP
Division of Nephrology, University of Arkansas for Medical Sciences, Little Rock, USA.
Singh
A B
AB
Shah
S V
SV
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D020170:Caspase 1; D053143:Caspase 2; D053148:Caspase 3; D053178:Caspase 6; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007672:Kidney Cortex / Q000502:physiology*; D008969:Molecular Sequence Data; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D011480:Protease Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000235:genetics; D051381:Rats; D015427:Reperfusion Injury / Q000235:genetics*
D018076:DNA, Complementary; D011480:Protease Inhibitors; D011506:Proteins; C505407:CASP3 protein, human; C505514:CASP6 protein, human; C505410:Casp3 protein, rat; C505518:Casp6 protein, rat; D053143:Caspase 2; D053148:Caspase 3; D053178:Caspase 6; D020169:Caspases; D003546:Cysteine Endopeptidases; D020170:Caspase 1