Proteinases participating in the processing and activation of prolegumain in primary cultured rat macrophages

2004 ◽  
Vol 385 (6) ◽  
pp. 511-516 ◽  
Author(s):  
F. Lecaille ◽  
D. Muno ◽  
E. Kominami ◽  
K. Ishidoh
Keyword(s):  
Cathepsin B ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Vacuolar Atpase ◽  
Cysteine Proteinases ◽  
Bafilomycin A1 ◽  
Atpase Inhibitor ◽  
Peptide Bonds ◽  
Mature Form

Abstract The mammalian legumain is a recently identified lysosomal cysteine proteinase belonging to the clan CD and homologous to plant legumain. This enzyme has the characteristic of specifically hydrolyzing peptide bonds after asparagine residues. As in the case of papain-type cysteine proteinases, legumain is synthesized as an inactive zymogen, and processed into a mature form localized in lysosomes. However, the mechanism of its activation remains unclear. In this study, we analyze which types of proteinases may participate in the processing of legumain in rat primary cultured macrophages using various proteinase inhibitors after 24 h treatment with Bafilomycin A1, a vacuolar ATPase inhibitor. The processing of legumain in macrophages was accomplished by papain-type cysteine proteinases other than cathepsin B.

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

2002 ◽  
Vol 383 (5) ◽  
pp. 839-842 ◽  
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.

scholarly journals Cystatins — Inhibitors of Cysteine Proteinases

1992 ◽  
Vol 3 (4) ◽  
pp. 307-332 ◽  
Author(s):  
Libuse A. Bobek ◽  
Michael J. Levine
Keyword(s):  
Common Ancestor ◽  
Molecular Level ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Diverse Group ◽  
Cysteine Proteinases ◽  
Future Directions ◽  
Cysteine Proteinase Inhibitors ◽  
Physicochemical And Functional Properties

The cystatin superfamily of proteins, derived from a common ancestor, is comprised of a diverse group of potent cysteine proteinase inhibitors and antibacterial/viral agents grouped into several families. This review concentrates on family 2 cystatins, namely, the human salivary cystatins and cystatin C. Emphasis is given to their physicochemical and functional properties at both the protein and the molecular level. The role of cystatins in disease processes, including those in the oral cavity, is also discussed. Finally, future directions for cystatin research in oral biology are presented.

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

2003 ◽  
Vol 77 (1) ◽  
pp. 21-26 ◽  
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.

scholarly journals Changes in Proteolytic Activity and Cysteine Proteinase Gene Expression during the Senescence of etr1-1 Transgenic Petunias

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1105B-1105
Author(s):  
Michelle Jones ◽  
Gunching Chaffin ◽  
David Clark
Keyword(s):  
Proteolytic Activity ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Transcript Abundance ◽  
Flower Senescence ◽  
Cysteine Proteinases ◽  
University Of Florida ◽  
Nitrogen Levels ◽  
Protein Levels ◽  
The University

Corolla senescence in petunias was accompanied by a decrease in total proteins and a corresponding increase in proteolytic activity. Transgenic petunias that contain the mutated ethylene receptor (35S:etr1-1) have reduced sensitivity to ethylene and delayed flower senescence. Declines in total protein levels and increases in proteolytic activity were also delayed in etr1-1 flowers and corresponded with corolla wilting. Experiments using class-specific proteinase inhibitors indicated that proteolytic activity in petunia corollas was largely due to cysteine proteinases. Total nitrogen levels within the corollas of both wild type and etr1-1 flowers also decreased during senescence. Nine cDNAs encoding putative cysteine proteinases (CPs) were identified from a petunia EST database developed at the University of Florida. Six of these cysteine proteinases showed increased transcript abundance during corolla senescence (senescence-associated CPs) while three decreased in abundance. Of the six senescence-associated cysteine proteinases, only five showed delayed up regulation in etr1-1 flowers that corresponded with corolla wilting. The role of ethylene in the regulation of protein degradation during flower senescence will be discussed.

Cytochemical and biochemical evidence of cathepsin B in malignant, transformed and normal breast epithelial cells

1987 ◽  
Vol 87 (1) ◽  
pp. 145-154
Author(s):  
E. Krepela ◽  
J. Bartek ◽  
D. Skalkova ◽  
J. Vicar ◽  
D. Rasnick ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cathepsin B ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Normal Breast ◽  
Breast Cancer Cell Lines ◽  
Human Breast ◽  
Breast Epithelial Cells ◽  
Breast Epithelial ◽  
Normal Breast Epithelial Cells

Human breast cancer cell lines, as well as transformed mammary epithelial cells (HBL-100) and growth-stimulated normal breast epithelial cells showed positive cytochemical reaction with the proteinase substrate 2-(N-benzyloxycarbonyl-L-arginyl-L-arginylamido)-4-methoxynapht halene, in the presence of 5-nitrosalicylaldehyde. The reaction product, small fluorescent granules, was distributed throughout the cytoplasm, in the perinuclear zone, in some cytoplasmic projections, and at the cell surface. Using a panel of various proteinase inhibitors, we found that the formation of the reaction product was an enzymic function of a cysteine proteinase. Using the substrate 7-(N-benzyloxycarbonyl-L-arginyl-L-arginylamido)-4-methylcoumarin, we evaluated some biochemical properties of the cysteine proteinase, including pH-activity profile, pH stability, apparent relative molecular mass and sensitivity toward various proteinase inhibitors. We found that the proteinase from the studied breast epithelial cells exhibited characteristics of a mature form of cathepsin B. Taken together, the cytochemical and biochemical data provide evidence that human breast epithelial cells of cancer origin, as well as in the transformed or growth-stimulated state express active cathepsin B and compartmentalize it into specific subcellular sites.

scholarly journals Identification of three stage-specific proteinases of Plasmodium falciparum.

1987 ◽  
Vol 166 (3) ◽  
pp. 816-821 ◽  
Author(s):  
P J Rosenthal ◽  
K Kim ◽  
J H McKerrow ◽  
J H Leech
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Specific Activity ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Serine Proteinase ◽  
Cysteine Proteinases ◽  
Malaria Parasites ◽  
Neutral Ph ◽  
Cysteine Proteinase Inhibitors

We have identified and characterized three stage-specific proteinases of Plasmodium falciparum that are active at neutral pH. We analyzed ring-, trophozoite-, schizont-, and merozoite-stage parasites by gelatin substrate PAGE and characterized the identified proteinases with class-specific proteinase inhibitors. No proteinase activity was detected with rings. Trophozoites had a 28 kD proteinase that was inhibited by inhibitors of cysteine proteinases. Mature schizonts had a 35-40 kD proteinase that also was inhibited by cysteine proteinase inhibitors. Merozoite fractions had a 75 kD proteinase that was inhibited by serine proteinase inhibitors. The stage-specific activity of these proteinases and the correlation between the effects of proteinase inhibitors on the isolated enzymes with the effects of the inhibitors on whole parasites suggest potential critical functions for these proteinases in the life cycle of malaria parasites.

scholarly journals Activation of procathepsin B in human hepatoma cells: the conversion into the mature enzyme relies on the action of cathepsin B itself

1993 ◽  
Vol 293 (2) ◽  
pp. 437-442 ◽  
Author(s):  
L Mach ◽  
H Schwihla ◽  
K Stüwe ◽  
A D Rowan ◽  
J S Mort ◽  
...  
Keyword(s):  
Cathepsin B ◽  
Mammalian Cells ◽  
Proteinase Inhibitors ◽  
Cysteine Proteinase ◽  
Human Hepatoma ◽  
Single Chain ◽  
Human Hepatoma Cells ◽  
Chain Form

In order to elucidate the processing mechanism of the lysosomal cysteine proteinase, cathepsin B, in mammalian cells, recombinant rat and human cathepsin B precursors were expressed in Saccharomyces cerevisiae. The active-site cysteine residue was changed to serine to prevent autoprocessing. When the purified proenzymes were incubated with the soluble fraction of postnuclear organelles obtained from human hepatoma HepG2 cells, processing to a 33 kDa form corresponding to the mature endogenous single-chain enzyme was observed. Inhibitors of metallo-, serine and aspartic proteinases exerted no significant effect on procathepsin B processing in vitro. However, the processing activity was effectively blocked by cysteine proteinase inhibitors, in particular E-64 and its cathepsin-B-selective derivative CA-074. Processing positions were identified by using anti-peptide antibodies specific for epitopes in the N- and C-terminal cleavage regions. The single-chain form produced in vitro was thus shown to contain an N-terminal extension of at least four residues relative to the mature lysosomal enzyme, as well as a C-terminal extension present in the proenzyme but usually absent in fully processed cathepsin B. On expression of the wild-type proenzyme in yeast, procathepsin B undergoes autoprocessing, yielding a single-chain form of the active enzyme, which contains similar N- and C-terminal extensions. These results indicate that maturation of procathepsin B in vivo in mammalian tissues relies on the proteolytic activity of cathepsin B itself.

scholarly journals The synthesis and properties of peptidylmethylsulphonium salts with two cationic residues as potential inhibitors of prohormone processing

1988 ◽  
Vol 256 (3) ◽  
pp. 989-994 ◽  
Author(s):  
A Zumbrunn ◽  
S Stone ◽  
E Shaw
Keyword(s):  
Cathepsin B ◽  
Proteinase Inhibitors ◽  
The Other ◽  
Serine Proteinases ◽  
Cysteine Proteinases ◽  
C Terminus ◽  
Prohormone Processing ◽  
Affinity Labelling ◽  
Potential Inhibitors ◽  
Cationic Residues

Peptidylmethylsulphonium salts incorporating consecutive basic residues at the C-terminus of the peptidyl portion such as -Arg-Arg-, -Arg-Lys-, -Lys-Lys- and -Lys-Arg- were synthesized and examined as proteinase inhibitors. Serine proteinases with a specificity directed towards hydrolysis at cationic residues were found to be unaffected by these derivatives. On the other hand, cysteine proteinases, cathepsin B and, in particular, clostripain were readily inactivated by affinity labelling. The reagents thus are of promise for the study of prohormone processing promoted by cysteine proteinases.

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