TY - JOUR
T1 - Purification and characterization of putative endothelin converting enzyme in bovine adrenal medulla
T2 - Evidence for a cathepsin D-like enzyme
AU - Sawamura, Tatsuya
AU - Kimura, Sadao
AU - Shinmi, Osamu
AU - Sugita, Yoshiki
AU - Yanagisawa, Masashi
AU - Goto, Katsutoshi
AU - Masaki, Tomoh
N1 - Funding Information:
This work was supported in part by grants from University of Tsukuba Special Project Research on Metabolism, the Ministries of Education, Science and Culture of Japan and Japan Ciba Geigy Foundation.
PY - 1990/5/16
Y1 - 1990/5/16
N2 - A specific and sensitive assay has been established for measurement of endothelin converting activity in a tissue extract. This assay is based on measuring endothelin-1 generated from big endothelin-1 by endothelin converting enzyme (ECE) with radioimmunoassay using an endothelin C-terminal specific antibody. By using this assay, we purified and characterized ECE in bovineadrenomedullary chromaffin granules. ECE was purified over 3,000 times by a combination of DEAE, hydrophobic and gel filtration chromatography. A molecular weight of ECE was estimated to be approximately 30,000 by gel filtration. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that ECE had three major components with estimated molecular weights of 45,000, 30,000 and 15,000 like bovine spleen cathepsin D. ECE had a pH optimum at 3.5 and was inhibited by pepstatin. These results strongly suggest that ECE is a cathepsin D-like aspartic protease.
AB - A specific and sensitive assay has been established for measurement of endothelin converting activity in a tissue extract. This assay is based on measuring endothelin-1 generated from big endothelin-1 by endothelin converting enzyme (ECE) with radioimmunoassay using an endothelin C-terminal specific antibody. By using this assay, we purified and characterized ECE in bovineadrenomedullary chromaffin granules. ECE was purified over 3,000 times by a combination of DEAE, hydrophobic and gel filtration chromatography. A molecular weight of ECE was estimated to be approximately 30,000 by gel filtration. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that ECE had three major components with estimated molecular weights of 45,000, 30,000 and 15,000 like bovine spleen cathepsin D. ECE had a pH optimum at 3.5 and was inhibited by pepstatin. These results strongly suggest that ECE is a cathepsin D-like aspartic protease.
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U2 - 10.1016/0006-291X(90)91160-T
DO - 10.1016/0006-291X(90)91160-T
M3 - Article
C2 - 2189405
AN - SCOPUS:0025280665
VL - 168
SP - 1230
EP - 1236
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 3
ER -