Interaction of human erythrocyte multicatalytic proteinase with polycations.
The multicatalytic proteinase from human erythrocytes (macropain, proteasome) is a large enzyme composed of at least six distinct subunits ranging in molecular masses from 20 to 30 kDa. As its name implies, this proteinase appears to contain multiple catalytic sites with differing specificities toward peptide substrates. Several polycationic substances, including polylysines, polyarginine, protamine and histone H1 markedly stimulated caseinolytic activity of the proteinase. Activation was instantaneous, and involved increasing the Vmax of the proteinase for casein. Prolonged preincubation with polylysine at 37 degrees C resulted in autolytic inactivation of the proteinase. The polylysine concentrations required for half-maximal activation or autolytic inactivation were the same. A 23 kDa subunit of the proteinase disappeared at the same rate as loss of catalytic activity, and with the same pH dependence and polylysine concentration dependence. These results suggest that polylysine perturbs the structure of the multicatalytic proteinase, resulting in increased catalytic activity toward substrates; and, with prolonged exposure, allowing autoproteolytic inactivation to occur. The 23 kDa subunit appeared to be required for expression of caseinolytic activity, and may therefore be a catalytic subunit of the complex having activity against casein.
Mellgren
R L
RL
Department of Pharmacology and Therapeutics, Medical College of Ohio, Toledo 43699-0008.
D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003546:Cysteine Endopeptidases / Q000097:blood* / Q000302:isolation & purification; D004789:Enzyme Activation; D004912:Erythrocytes / Q000201:enzymology*; D006657:Histones / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000097:blood* / Q000302:isolation & purification; D010455:Peptides / Q000494:pharmacology; D011107:Polylysine / Q000494:pharmacology; D011479:Protamines / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D013096:Spermine / Q000494:pharmacology
D006657:Histones; D009097:Multienzyme Complexes; D010455:Peptides; D011479:Protamines; D011480:Protease Inhibitors; D011107:Polylysine; C015462:polyarginine; D013096:Spermine; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex