In vitro
 and in vivo
 antifungal properties of cysteine proteinase inhibitor from green kiwifruit

Expression of the 6-hydroxy-D-nicotine oxidase (6-HDNO) gene from Arthrobacter oxidans cloned into Escherichia coli showed a marked temperature-dependence. Transformed E. coli cells grown at 30 degrees C exhibited a several-fold higher 6-HDNO activity than did cells grown at 37 degrees C. This effect did not depend on the promoter used for expression of the cloned gene in E. coli, nor was it an effect of 6-HDNO mRNA instability at 37 degrees C. Studies performed in vivo and in vitro revealed that an increased susceptibility of apo-6-HDNO to proteolytic attack at 37 degrees C was responsible for the observed phenomenon. Extracts from cells grown at 37 degrees C showed on Western blots a decrease in immunologically detectable 6-HDNO polypeptide when compared with extracts from cells grown at 30 degrees C. The 6-HDNO polypeptide is covalently modified by attachment of the cofactor FAD to a histidine residue. It could be shown that covalent flavinylation of the apoenzyme in vitro, i.e. formation of holoenzyme, by incubation of cell extracts with FAD and phosphoenolpyruvate protected the 6-HDNO polypeptide from degradation at 37 degrees C. Of a variety of proteinase inhibitors tested only the cysteine-proteinase inhibitor L-3-trans-carboxyoxiran-2-carbonyl-L-leucylagmatine (E64) prevented degradation, by up to 70%, of the apoenzyme.

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Involvement of cysteine proteinases in excystment of Paragonimus ohirai metacercariae induced by sodium cholate and A23187

Journal of Helminthology ◽

10.1079/joh2002144 ◽

2003 ◽

Vol 77 (1) ◽

pp. 21-26 ◽

Cited By ~ 8

Author(s):

T. Ikeda

Keyword(s):

Proteinase Inhibitor ◽

Proteinase Inhibitors ◽

Cysteine Proteinase ◽

Serine Proteinase ◽

Sodium Cholate ◽

Cysteine Proteinase Inhibitor ◽

Cysteine Proteinases ◽

Serine Proteinase Inhibitor ◽

Similar Activity

AbstractThe involvement of intrinsic proteinases in the excystment of Paragonimus ohirai metacercariae was studied in in vitro excystment induced by sodium (Na) cholate, a bile salt and A23187, a Ca2+ ionophore. The effects of various proteinase inhibitors on the in vitro excystment were examined and similar inhibitory profiles were obtained. Benzyloxycarbonyl-L-leucyl-L-leucinal (Z-Leu-Leu-H), a cysteine proteinase inhibitor and 4-(2-aminoethyl)-benzenesulfonyl fluoride (Pefabloc SC), a serine proteinase inhibitor completely inhibited excystment, while L-3-carboxy-2,3-trans-epoxypropionyl-leucylamido (4-guanidino)-butane (E-64), a cysteine proteinase inhibitor and leupeptin, a cysteine/serine proteinase inhibitor permitted partial excystment at a lower rate, but inhibited it from proceeding from the partial excystment stage. In secretions released from metacercariae during excystment, proteinase activities detected towards various fluorogenic peptidyl substrates were almost completely inhibited by Z-Leu-Leu-H and E-64, but not by Pefabloc SC. Sodium cholate induced a higher secretion of cysteine proteinases and a higher rate of excystment than A23187. Profiles of cysteine proteinase activities towards five peptidyl substrates detected were markedly different among the two secretions and the lysate of newly excysted juveniles. Newly excysted juveniles released cysteine proteinases with similar activity profiles and levels to metacercariae induced by Na cholate-incubation, whereas the release of cysteine proteinases was reduced compared with metacercariae induced by A23187-incubation. These results provide valuable information about the involvement of intrinsic proteinases in metacercarial excystment.

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Proteolytically active complexes of cathepsin L and a cysteine proteinase inhibitor; purification and demonstration of their formation in vitro

Archives of Biochemistry and Biophysics ◽

10.1016/0003-9861(92)90734-e ◽

1992 ◽

Vol 294 (2) ◽

pp. 623-629 ◽

Cited By ~ 23

Author(s):

Robert N. Pike ◽

Theresa H.T. Coetzer ◽

Clive Dennison

Keyword(s):

Proteinase Inhibitor ◽

Cysteine Proteinase ◽

Cathepsin L ◽

Cysteine Proteinase Inhibitor

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Inhibition of human liver cathepsin L by α2 cysteine-proteinase inhibitor and the low-Mr cysteine proteinase inhibitor from human serum

Biochemical Journal ◽

10.1042/bj2200147 ◽

1984 ◽

Vol 220 (1) ◽

pp. 147-155 ◽

Cited By ~ 12

Author(s):

M Pagano ◽

F Esnard ◽

R Engler ◽

F Gauthier

Keyword(s):

Human Serum ◽

Proteinase Inhibitor ◽

Human Liver ◽

Physiological Function ◽

Proteinase Inhibitors ◽

Cysteine Proteinase ◽

Cathepsin L ◽

Cysteine Proteinase Inhibitor ◽

Lysosomal Proteinases

The inhibition of human liver cathepsin L by two specific proteinase inhibitors present in human serum, namely alpha 2 cysteine-proteinase inhibitor and the low-Mr cysteine-proteinase inhibitor, was studied. Kinetic parameters, including inhibition constants (Ki) and rate constants for association and dissociation (k+1 and K-1), were determined. The values found are consistent with a possible physiological function of these inhibitors to control cathepsin L activity. Furthermore, a transfer of active proteinase from the complex with either cysteine-proteinase inhibitor species to alpha 2-macroglobulin was demonstrated in vitro. Given the rate of dissociation of both cathepsin-L-cysteine-proteinase inhibitor complexes, a function of transitory inhibitor can therefore be hypothesized for these proteins and might then provide an explanation of the clearance of lysosomal proteinases.

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Effect of Cysteine Proteinase Inhibitors on Murine B16 Melanoma Cell Invasion in vitro

Biological Chemistry ◽

10.1515/bc.2002.088 ◽

2002 ◽

Vol 383 (5) ◽

pp. 839-842 ◽

Cited By ~ 24

Author(s):

Natasa Sever ◽

Metka Filipic ◽

Joze Brzin ◽

Tamara T. Lah

Keyword(s):

Cell Invasion ◽

Cathepsin B ◽

Proteinase Inhibitor ◽

Proteinase Inhibitors ◽

Cysteine Proteinase ◽

Cathepsin L ◽

Cysteine Proteinase Inhibitor ◽

Cysteine Proteinases ◽

Cysteine Proteinase Inhibitors

Abstract Various types of proteinases are implicated in the malignant progression of human and animal tumors. Proteinase inhibitors may therefore be useful as therapeutic agents in antiinvasive and antimetastatic treatment. The aims of this study were (1) to estimate the relative importance of proteinases in B16 cell invasion in vitro using synthetic, classspecific proteinase inhibitors and (2) to assess the inhibitory effect of some naturally occurring cysteine proteinase inhibitors. Serine proteinase inhibitor reduced invasiveness by up to 24%, whereas inhibition of aspartic proteinases reduced invasion by 11%. Synthetic inhibitors of cysteine proteinases markedly impaired invasion: cathepsin B inhibitors, particularly Ca 074Me, inhibited invasion from 20 40%, whereas cathepsin L inhibitor Clik 148 reduced invasion by 11%. The potato cysteine proteinase inhibitor PCPI 8.7 inhibited invasion by 21%, whereas another potato inhibitor, PCPI 6.6, and the mushroom cysteine proteinase inhibitor clitocypin had no effects. As the inhibitors that inhibited cathepsin B were in general more efficient at impairing the invasiveness, we conclude that of the two cysteine proteinases, cathepsin B plays a more important role than cathepsin L in murine melanoma cell invasion.

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