Cathepsin S The cysteine proteinase from bovine lymphoid tissue is distinct from cathepsin L (EC 3.4.22.15)

Cathepsin S was purified from bovine spleen by acid autolysis, (NH4)2SO4 fractionation and chromatography on CM-Sephadex C-50, CM-cellulose and activated-thiol-Sepharose. Cathepsin L was isolated from lysosomal fractions of rat liver, rat kidney and bovine liver. Generally, cathepsin L was bound tightly to CM-Sephadex C-50. Preparations of cathepsin L from rat liver, rat kidney and bovine liver were shown to have kinetic constants for the substrate benzyloxycarbonyl-Phe-Arg-7-(4-methyl)coumarylamide in the same range (Km 2-3 microM). Benzyloxycarbonyl-Phe-Phe-diazomethane proved to be a sensitive irreversible inhibitor of cathepsin L from different species. Cathepsin S differed in all these characteristics from cathepsin L. A polyclonal antibody to cathepsin L from rat reacted with bovine cathepsin L but not with bovine cathepsin S.

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Cathepsin S from bovine spleen. Purification, distribution, intracellular localization and action on proteins

Biochemical Journal ◽

10.1042/bj2640467 ◽

1989 ◽

Vol 264 (2) ◽

pp. 467-473 ◽

Cited By ~ 142

Author(s):

H Kirschke ◽

B Wiederanders ◽

D Brömme ◽

A Rinne

Keyword(s):

Cathepsin L ◽

Intracellular Localization ◽

Gel Filtration ◽

Similar Rate ◽

Cathepsin S ◽

Purification Method ◽

Cytoplasmic Granules ◽

Bovine Kidney ◽

Ph Values ◽

C 50

Cathepsin S was detected in bovine kidney, spleen, lymph nodes and lung by immunochemical methods. The immunostaining of cathepsin S in kidney was concentrated to the cells of the proximal tubule, where the enzyme was present in cytoplasmic granules. The purification method for cathepsin S from bovine spleen involved (NH4)2SO4 fractionation, chromatography on CM-Sephadex C-50, gel filtration on Sephacryl S-200 and chromatofocusing (pH 8.0-6.0). The enzyme was partially destroyed by autolysis of the homogenate at pH 4.2. The isoelectric point of cathepsin S was 7.0. Cathepsin S was found to hydrolyse proteins at a similar rate to cathepsin L below pH 7.0. At pH values of 7.0-7.5 cathepsin S retained most of its activity, whereas cathepsin L was completely inactive.

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The specificity of bovine spleen cathepsin S. A comparison with rat liver cathepsins L and B

Biochemical Journal ◽

10.1042/bj2640475 ◽

1989 ◽

Vol 264 (2) ◽

pp. 475-481 ◽

Cited By ~ 96

Author(s):

D Brömme ◽

A Steinert ◽

S Friebe ◽

S Fittkau ◽

B Wiederanders ◽

...

Keyword(s):

Rat Liver ◽

Cathepsin L ◽

Peptide Bond ◽

Cathepsin S ◽

Cleavage Sites ◽

Amino Acid Residues ◽

Peptide Substrates ◽

Hydrophobic Residues ◽

Micromolar Range ◽

Better Than

The peptide-bond-specificity of bovine spleen cathepsin S in the cleavage of the oxidized insulin B-chain and peptide methylcoumarylamide substrates was investigated and the results are compared with those obtained with rat liver cathepsins L and B. Major cleavage sites in the oxidized insulin B-chain generated by cathepsin S are the bonds Glu13-Ala14, Leu17-Val18 and Phe23-Tyr26; minor cleavage sites are the bonds Asn3-Gln4, Ser9-His10 and Leu15-Tyr16. The bond-specificity of this proteinase is in part similar to the specificities of cathepsin L and cathepsin N. Larger differences are discernible in the reaction with synthetic peptide substrates. Cathepsin S prefers smaller neutral amino acid residues in the subsites S2 and S3, whereas cathepsin L efficiently hydrolyses substrates with bulky hydrophobic residues in the P2 and P3 positions. The results obtained from inhibitor studies differ somewhat from those based on substrates. Z-Phe-Ala-CH2F (where Z- represents benzyloxycarbonyl-) is a very potent time-dependent inhibitor for cathepsin S, and inhibits this proteinase 30 times more efficiently than it does cathepsin L and about 300 times better than it does cathepsin B. By contrast, the peptidylmethanes Z-Val-Phe-CH3 and Z-Phe-Lys(Z)-CH3 inhibit competitively both cathepsin S and cathepsin L in the micromolar range.

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Identification of bile acid-CoA: amino acid N-acyltransferase in rat kidney

Biochemical Journal ◽

10.1042/bj2800821 ◽

1991 ◽

Vol 280 (3) ◽

pp. 821-824 ◽

Cited By ~ 8

Author(s):

J B Kwakye ◽

M R Johnson ◽

S Barnes ◽

W E Grizzle ◽

R B Diasio

Keyword(s):

Bile Acid ◽

Amino Acid ◽

Molecular Mass ◽

Rat Liver ◽

Polyclonal Antibody ◽

Human Liver ◽

Acid Metabolism ◽

Rat Kidney ◽

Immunoblot Analysis ◽

Bile Acid Metabolism

A novel location of the bile-acid-conjugating enzyme bile acid-CoA:amino acid N-acyltransferase (BAT) has been discovered in the cytosolic fraction of rat kidney. Both taurine and glycine were utilized as substrates. Formation of bile acid N-acyl amidates was verified by h.p.l.c. by comparison with authentic standards and by specific hydrolysis using cholylglycine hydrolase. Immunoblot analysis using a human liver anti-BAT polyclonal antibody indicated that rat kidney BAT has the same molecular mass as rat liver BAT. These findings suggest that the kidney has a role in bile acid metabolism and physiology.

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Rat kidney carboxylesterase. Cloning, sequencing, cellular localization, and relationship to rat liver hydrolase.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)43935-x ◽

1994 ◽

Vol 269 (47) ◽

pp. 29688-29696

Author(s):

B Yan ◽

D Yang ◽

M Brady ◽

A Parkinson

Keyword(s):

Rat Liver ◽

Cellular Localization ◽

Rat Kidney

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Differential Regulation of Cathepsin S and Cathepsin L in Interferon γ–treated Macrophages

Journal of Experimental Medicine ◽

10.1084/jem.20020978 ◽

2003 ◽

Vol 197 (2) ◽

pp. 169-179 ◽

Cited By ~ 79

Author(s):

Courtney Beers ◽

Karen Honey ◽

Susan Fink ◽

Katherine Forbush ◽

Alexander Rudensky

Keyword(s):

Cathepsin L ◽

Specific Inhibitor ◽

Cell Types ◽

Interferon Γ ◽

Cathepsin S ◽

Antigen Presenting Cell ◽

Relative Contribution ◽

Helper Cell Type ◽

Ifn Γ ◽

Antigen Presenting

Cathepsin S (catS) and cathepsin L (catL) mediate late stages of invariant chain (Ii) degradation in discrete antigen-presenting cell types. Macrophages (Mϕs) are unique in that they express both proteases and here we sought to determine the relative contribution of each enzyme. We observe that catL plays no significant role in Ii cleavage in interferon (IFN)-γ–stimulated Mϕs. In addition, our studies show that the level of catL activity is significantly decreased in Mϕs cultured in the presence of IFN-γ whereas catS activity increases. The decrease in catL activity upon cytokine treatment occurs despite the persistence of high levels of mature catL protein, suggesting that a specific inhibitor of the enzyme is up-regulated in IFN-γ–stimulated peritoneal Mϕs. Similar inhibition of activity is observed in dendritic cells engineered to overexpress catL. Such enzymatic inhibition in Mϕs exhibits only partial dependence upon Ii and therefore, other mechanisms of catL inhibition are regulated by IFN-γ. Thus, during a T helper cell type 1 immune response catL inhibition in Mϕs results in preferential usage of catS, such that major histocompatibility complex class II presentation by all bone marrow–derived antigen-presenting cell is regulated by catS.

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The role of the cysteine proteinase cathepsin S in astrocytoma invasion

Neuropathology and Applied Neurobiology ◽

10.1046/j.1365-2990.2002.39286_23.x ◽

2002 ◽

Vol 28 (2) ◽

pp. 156-156

Author(s):

D. Gibson ◽

T. Flannery ◽

K. Mulligan ◽

M. Mirakhur ◽

D. McCormick

Keyword(s):

Cysteine Proteinase ◽

Cathepsin S

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Squamous cell carcinoma antigen is a potent inhibitor of cysteine proteinase cathepsin L

FEBS Letters ◽

10.1016/0014-5793(94)01456-b ◽

1995 ◽

Vol 359 (1) ◽

pp. 78-80 ◽

Cited By ~ 38

Author(s):

Atsushi Takeda ◽

Takako Yamamoto ◽

Yoshiko Nakamura ◽

Tadahito Takahashi ◽

Toshihiko Hibino

Keyword(s):

Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Squamous Cell ◽

Potent Inhibitor ◽

Cysteine Proteinase ◽

Cathepsin L ◽

Squamous Cell Carcinoma Antigen

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Cleavage of immunoglobulin G by excretory–secretory cathepsin S-like protease of Spirometra mansoni plerocercoid

Parasitology ◽

10.1017/s0031182000076496 ◽

1994 ◽

Vol 109 (5) ◽

pp. 611-621 ◽

Cited By ~ 56

Author(s):

Y. Kong ◽

Y.-B. Chung ◽

S.-Y. Cho ◽

S.-Y. Kang

Keyword(s):

Immunoglobulin G ◽

Cathepsin L ◽

Immunoblot Analysis ◽

Cathepsin S ◽

Terminal Amino Acid ◽

Optimal Activity ◽

Spirometra Mansoni ◽

Terminal Amino ◽

Secretory Products ◽

Ph Range

When immunoglobulin G (IgG) was incubated with Spirometra mansoni plerocercoid (sparganum), it was cleaved into Fab and Fc fragments. Fab/c fragments were also hydrolysed. The digestion was accelerated by dithiothreitol (DTT), indicating that cleavage of IgG heavy chain was due to a cysteine protease secreted into the medium. The responsible enzyme, of Mr 27 (± 0·8) kDa, was purified by a series of thiopropyl affinity, Sephacryl S-300 HR and DEAE-anion exchange chromatographies, either from worm extracts or from excretory–secretory products (ESP). The purified, thiol-dependent protease showed an optimal activity at pH 5·7 with 0·1 M sodium acetate but was active over the pH range 4·5–8·0. Its activity was inhibited completely by 10−5 M L-trans-epoxysuccinylleucylamido(4-guanidino) butane (E-64) and 1 mM iodoacetamide (IAA), but by only 53% using the specific cathepsin L inhibitor, Z-Phe-Phe-CHN2 (5 × 10−5 M). Partial NH2-terminal amino acid sequence was Leu-Pro-Asp-Ser-Val-Asn-Trp-Arg-Glu-Gly-Ala-Val-Thr-Ala-Val which showed 80% homology to human cathepsin S. Immunoblot analysis showed that sera from infected patients exhibited IgE antibody reaction. It is proposed that cleavage of immunoglobulin by an excreted–secreted, cathepsin S-like, allergenic protease is a mechanism of immune evasion used by the sparganum.

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