SUBCELLULAR-LOCALIZATION, PARTIAL-PURIFICATION, AND CHARACTERIZATION OF A DYNORPHIN PROCESSING ENDOPEPTIDASE FROM BOVINE PITUITARY

被引:27
作者
DEVI, L [1 ]
GUPTA, P [1 ]
FRICKER, LD [1 ]
机构
[1] YESHIVA UNIV ALBERT EINSTEIN COLL MED,DEPT MOLEC PHARMACOL,BRONX,NY 10461
关键词
NEUROPEPTIDE BIOSYNTHESIS; TRYPSIN-LIKE; OPIOID PEPTIDE; ENKEPHALIN; ENDORPHIN;
D O I
10.1111/j.1471-4159.1991.tb02598.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
An enzyme capable of cleaving dynorphin B-29 to dynorphin B-13 is present in bovine pituitary, with 40- to 50-fold higher specific activity in the posterior and intermediate lobes than in the anterior lobe. Subcellular fractionation of bovine neurointermediate pituitary shows that this enzyme is present in the peptide-containing secretory vesicles. The enzyme has been purified 2,800-fold from whole bovine pituitaries using ion-exchange and gel filtration chromatography. Purified dynorphin-converting enzyme has a neutral pH optimum, and is substantially inhibited by the thiol-protease inhibitor p-chloromercuriphenylsulfonic acid, but not by serine or metalloprotease inhibitors. The purified enzyme processes dynorphin B-29 at Arg14, producing both dynorphin B-14 and dynorphin B-13 in a 5:1 ratio. No other cleavages are observed, suggesting that the activity is free from other proteases and is specific for single Arg sequences. Purified enzyme also processes dynorphin A-17 at the single Arg cleavage site, generating both dynorphin A-8 and A-9 in a 7:1 ratio. The tissue distribution, subcellular localization, and substrate specificity of this enzyme are consistent with a physiological role in the processing of dynorphin B-29 and dynorphin A-17, and possibly other peptides, at single Arg residues.
引用
收藏
页码:320 / 329
页数:10
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