The generation of big-endothelin (1-22) (endothelin-valine) from big-endothelin in the soluble fraction of porcine lung.
The degradation of big-endothelin (big-ET) in the soluble fraction of porcine lung was investigated. The degradation in the presence of p-chloromercuribenzoate (PCMB), pepstatin A, and EDTA resulted in the accumulation of two newly-formed fragments, big-ET (23-39) and big-ET (1-22), the latter called endothelin-valine (ET-Val). The generation of the two fragments was inhibited by diisopropylfluorophosphate (DFP). The enzyme responsible, called ET-Val-generating endopeptidase, was isolated from porcine lung by a procedure including chromatographies on columns of DEAE-cellulose, hydroxylapatite, Mono Q, p-mercuribenzoate-Sepharose, and Superose 6. The molecular weight of the enzyme was 140,000 and the pH optimum of the activity was 7.0. The activity was strongly inhibited by DFP, but scarcely inhibited by PCMB, EDTA, and pepstatin A. Thus, the isolated enzyme was classified as a serine protease cleaving big-ET at the Val22-Asn23 bond.
Watanabe
T
T
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Yokosawa
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002850:Chromatography, Gel; D019332:Endothelin-1; D016232:Endothelins / Q000737:chemistry / Q000378:metabolism*; D008168:Lung / Q000201:enzymology / Q000378:metabolism*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D012995:Solubility; D013552:Swine
D019332:Endothelin-1; D016232:Endothelins; D011498:Protein Precursors; C075556:big-endothelin (1-22); D012697:Serine Endopeptidases; C075555:endothelin-valine-generating endopeptidase