Identification of a novel ATP-dependent proteolytic activity in mitochondrial intermembrane space.
The formation of primary amines via proteolysis was monitored in isolated rat liver, kidney cortex and heart mitochondria in the presence and in the absence of ATP. The highest proteolytic activity was detected in kidney cortex mitochondria with about 120 nmoles primary amines/hour x mg protein. The formation rates of liver mitochondria amounted to about 100 nmoles primary amines/hour x mg protein and in heart mitochondria about 60 nmoles primary amines/hour x mg protein. In all mitochondria investigated an ATP-dependent proteolysis of 20-40 nmoles primary amines/hour x mg protein was detected. The effects of various protease inhibitors were tested in rat liver mitochondria and thiol-specific reagents showed a 35-70% inhibition. The ATP stimulable portion of proteolysis was blocked by hemin, a known inhibitor of ATP-dependent proteases. The localization of the proteolytic activity was tested by fractionation of the compartments of rat liver mitochondria using the flourogenic peptide suc-Leu-Leu-Val-Tyr-MCA as substrate. About 90% of the ATP-dependent peptide cleavage activity were found in the mitochondrial intermembrane space. The characteristics of the enzyme were compared to those of other known mitochondrial ATP-dependent proteases and it was concluded that it represents a novel proteolytic system of the intermembrane space.
Sitte
N
N
Institute of Biochemistry, Medical Faculty Charité, Humboldt-University Berlin, Germany.
Drung
I
I
Dubiel
W
W
D049069:ATP-Dependent Proteases; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D006360:Heat-Shock Proteins / Q000378:metabolism*; D007425:Intracellular Membranes / Q000201:enzymology*; D007672:Kidney Cortex / Q000201:enzymology; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology*; D008929:Mitochondria, Heart / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology / Q000648:ultrastructure; D008969:Molecular Sequence Data; D051381:Rats; D017208:Rats, Wistar; D012697:Serine Endopeptidases / Q000378:metabolism*; D013347:Subcellular Fractions / Q000201:enzymology; D013367:Submitochondrial Particles / Q000201:enzymology*; D013379:Substrate Specificity
D006360:Heat-Shock Proteins; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases; C111985:mitochondrial intermembrane space protease; D000251:Adenosine Triphosphatases