Microbial serine carboxypeptidase inhibitors--comparative analysis of actions on homologous enzymes derived from man, yeast and wheat.
The actions of peptidase inhibitors derived from Streptomycete on human cathepsin A (hCath A), yeast carboxypeptidase Y (CPY), and wheat carboxypeptidase II (CPW) were analyzed comparatively. Lactacystin and omuralide (clasto-lactacystin beta-lactone), well-known cytoplasmic proteasome inhibitors, both had a potent and non-competitive inhibitory effect on these homologous serine carboxypeptidases, although they inhibited CPW and hCath A more effectively than CPY in vitro. Ebelactone B exhibited a mixed non-competitive inhibitory effect and selectivity for CPY. Piperastatin A showed competitive inhibition of CPY and hCath A but had little effect on CPW. In contrast, chymostatin inhibited CPW efficiently, while it had less effect on hCath A and CPY. In cell culture system, lactacystin was the most potent as to inactivation of the intralysosomal recombinant hCath A activity expressed in a genetically engineered fibroblastic cell line with galactosialidosis (hCath A deficiency). These results suggest that the specific inhibitory effects of lactacystin and its derivatives on hCath A might be applicable to elucidate the pathophysiological roles in the human deficinecy.
Division of Medicinal Biotechnology, Institute for Medicinal Resources, Graduate School of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770-8505, Japan.
Yurie
Y
Satoh
Yoshito
Y
Kadota
Yukako
Y
Oheda
Jun
J
Kuwahara
Seiichi
S
Aikawa
Fumiko
F
Matsuzawa
Hirofumi
H
Doi
Takaaki
T
Aoyagi
Hitoshi
H
Sakuraba
Kohji
K
Itoh
D002268:Carboxypeptidases / Q000037:antagonists & inhibitors*; D043402:Cathepsin A / Q000037:antagonists & inhibitors / Q000378:metabolism; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000378:metabolism; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D015151:Immunoblotting; D009097:Multienzyme Complexes / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D013045:Species Specificity; D013302:Streptomyces / Q000737:chemistry; D014908:Triticum / Q000201:enzymology*
D018076:DNA, Complementary; D009097:Multienzyme Complexes; D029701:Saccharomyces cerevisiae Proteins; D015842:Serine Proteinase Inhibitors; D002268:Carboxypeptidases; D043402:Cathepsin A; C499283:PRC1 protein, S cerevisiae; C022633:serine carboxypeptidase; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex