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.
Satoh
Yurie
Y
Division of Medicinal Biotechnology, Institute for Medicinal Resources, Graduate School of Pharmaceutical Sciences, The University of Tokushima, Tokushima 770-8505, Japan.
Kadota
Yoshito
Y
Oheda
Yukako
Y
Kuwahara
Jun
J
Aikawa
Seiichi
S
Matsuzawa
Fumiko
F
Doi
Hirofumi
H
Aoyagi
Takaaki
T
Sakuraba
Hitoshi
H
Itoh
Kohji
K
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