Affinity purification of endothia protease with a novel renin inhibitor SQ 32,970.
A novel tripeptidic renin inhibitor is described, SQ 32,970, that will potently inhibit endothia protease. This inhibitor can be coupled to Sepharose and will allow the affinity-purification of endothia protease in one step to greater than 95% purity as measured by SDS PAGE. The purified endothia protease cleaves the Lys-Pro-Ala-Glu-Phe-Nph-Arg-Leu substrate at the Phe-Nph bond with a Kcat/Km of 7445 (s-1 mM-1) at pH 3.1 and 4057 (s-1 mM-1) at pH 6.0. Affinity purified endothia protease can be crystallized in the pH range in which it is enzymatically active and can be inhibited by renin inhibitors.
Norman
J A
JA
Department of Pharmacology, Squibb Institute for Medical Research, Princeton, New Jersey.
Little
D
D
Free
C A
CA
Dejneka
T
T
Weber
H
H
Ryono
D E
DE
D001203:Ascomycota / Q000032:analysis*; D016282:Aspartic Acid Endopeptidases; D002851:Chromatography, High Pressure Liquid; D010450:Endopeptidases / Q000302:isolation & purification*; D005656:Fungal Proteins / Q000302:isolation & purification*; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D009842:Oligopeptides / Q000494:pharmacology*; D011480:Protease Inhibitors; D013379:Substrate Specificity; D000092502:Renin Inhibitors
D016282:Aspartic Acid Endopeptidases; D010450:Endopeptidases; D005656:Fungal Proteins; D009842:Oligopeptides; D011480:Protease Inhibitors; D000092502:Renin Inhibitors; C059981:SQ 32970; C042466:Endothia aspartic proteinase