Cathepsin L activity is increased in alveolar macrophages and bronchoalveolar lavage fluid of smokers.
AuthorsTakahashi H, Ishidoh K, Muno D, Ohwada A, Nukiwa T, Kominami E, Kira S
JournalAm Rev Respir Dis
PubMed ID8503570
'Elastinolytic enzymes derived from alveolar macrophages (AM) are considered to play an important role in the development of emphysema associated with cigarette smoking. In this study, the enzyme activity and mRNA expression of cathepsin L were quantitated in AM and bronchoalveolar lavage (BAL) fluid obtained from current smokers and compared ... More
Characterization of the serine protease and serine protease inhibitor from the tissue-penetrating nematode Anisakis simplex.
AuthorsMorris SR, Sakanari JA
JournalJ Biol Chem
PubMed ID7961683
'A serine protease and a serine protease inhibitor were purified from infective larvae of the parasitic nematode Anisakis simplex. The serine protease was found to be trypsin-like and preferentially cleaved substrates with the basic amino acid arginine at the P1 position (Z-Gly-Pro-Arg-AMC (where Z is benzyloxycarbonyl), Km = 0.019 mM, ... More
Changes of lysosomal proteinase activities and their expression in rat cultured keratinocytes during differentiation.
AuthorsTanabe H, Kumagai N, Tsukahara T, Ishiura S, Kominami E, Nishina H, Sugita H
JournalBiochim Biophys Acta
PubMed ID1911879
'The cathepsins B, H and L, lysosomal cysteine proteinases, play a major role in intracellular protein degradation. These proteinase activities and expressions were examined in a Ca2+ regulated epidermal culture system which consists of two morphological cell types: undifferentiated cells grown in low Ca2+ (0.1 mM concentration) and differentiated cells ... More
Alterations in cysteine proteinase content of rat lung associated with development of Pneumocystis carinii infection.
AuthorsHayes DJ, Stubberfield CR, McBride JD, Wilson DL
JournalInfect Immun
PubMed ID1894363
'The rate of hydrolysis of three cysteine-type proteinase substrates, N-benzyloxycarbonyl-Arg-Arg-4-methyl-7-coumarylamide (AMC) (cathepsin B), Arg-AMC (cathepsin H), and N-benzyloxycarbonyl-Phe-Arg-AMC (cathepsin L), were determined in rat lung throughout the time course of the induction of Pneumocystis carinii infection by immunosuppression. Cathepsin B-like and cathepsin L-like activities fell below control values initially, but ... More
Purification, characterization, and localization of follipsin, a novel serine proteinase from the fluid of porcine ovarian follicles.
AuthorsHamabata T, Okimura H, Yokoyama N, Takahashi T, Takahashi K
JournalJ Biol Chem
PubMed ID8027045
'Follipsin, an enzyme that accumulates in the follicular fluid of porcine ovaries during follicular maturation, was purified to apparent homogeneity. The purified enzyme consists of two different polypeptide chains having M(r) = 45,000 and 32,000 each, associated covalently. The enzyme activity was strongly inhibited by diisopropyl fluorophosphate, benzamidine, leupeptin, and ... More
Unique cleavage specificity of 'prohormone thiol protease' related to proenkephalin processing.
AuthorsAzaryan AV, Hook VY
JournalFEBS Lett
PubMed ID8137939
'''Prohormone thiol protease'' (PTP) represents the major enkephalin precursor processing activity in chromaffin granules. In this study, cleavage specificity of PTP for paired basic and monobasic residues was examined with a series of model peptide-MCA (-methylcoumarinamide) substrates. Monobasic peptides were cleaved at the COOH- and NH2-terminal sides of the single ... More
Action of rat liver cathepsin L on collagen and other substrates.
AuthorsKirschke H, Kembhavi AA, Bohley P, Barrett AJ
JournalBiochem J
PubMed ID7082295
'1. It has been found that cathepsin L is very susceptible to loss of activity through autolysis. When this is prevented by purification and storage of the enzyme as its mercury derivative, preparations are obtained with higher specific activity than previously. 2. Active-site titration shows, however, that even the new ... More
Engineering of papain: selective alteration of substrate specificity by site-directed mutagenesis.
AuthorsKhouri HE, Vernet T, Ménard R, Parlati F, Laflamme P, Tessier DC, Gour-Salin B, Thomas DY, Storer AC
JournalBiochemistry
PubMed ID1892810
'The S2 subsite specificity of the plant protease papain has been altered to resemble that of mammalian cathepsin B by site-directed mutagenesis. On the basis of amino acid sequence alignments for papain and cathepsin B, a double mutant (Val133Ala/Ser205Glu) was produced where Val133 and Ser205 are replaced by Ala and ... More
Purification and properties of a neutral thiol protease from larval trematode parasite Paragonimus westermani metacercariae.
AuthorsYamakami K, Hamajima F
JournalComp Biochem Physiol B
PubMed ID3304826
'1. A neutral thiol protease was isolated from the extract of larvae of the mammalian trematode parasite, Paragonimus westermani metacercariae, by arginine-Sepharose, Ultrogel AcA-54 and DEAE-toyopearl column chromatography, measuring its activity by the hydrolysis of Boc-Val-Leu-Lys-MCA as a substrate. 2. The molecular weight of the purified enzyme was estimated to ... More
Distribution of cathepsins B and L in the kidney and their role in tubular protein absorption.
AuthorsOlbricht CJ
JournalEur J Clin Chem Clin Biochem
PubMed ID1493160
'Intralysosomal proteinase activity, due to cathepsins B and L, was measured in microdissected segments of rat nephron, using Z-Phen-Arg-7-amido-4-methylcoumarin as the substrate. Cathepsin B was determined with Z-Arg-Arg-7-amido-4-methylcoumarin. The enzymes cleave on the carboxy side of arginine and release free 7-amino-4-methylcoumarin, which is highly fluorescent and can be measured at ... More
Human plasma kallikrein. A rapid purification method with high yield.
AuthorsNagase H, Barrett AJ
JournalBiochem J
PubMed ID6914195
'A simple method for isolation of kallikrein from human plasma is described. Before activation of the enzyme with acetone, the plasma was treated with 0.2 M-methylamine at pH 8.2 to inactivate alpha 2-macroglobulin and thus prevent the irreversible binding of the active enzyme to the inhibitor. The enzyme was adsorbed ... More
Characterization of an endopeptidase of Trypanosoma brucei brucei.
'A soluble 80-kDa endopeptidase has been isolated from Trypanosoma brucei brucei. The enzyme, which has a pI 5.1, is optimally active at about pH 8.2 and has apparent pKa values of 6.0 and greater than or equal to 10. It is inhibited by the serine protease inhibitor diisopropylfluorophosphate and by ... More
Brugia pahangi: identification and characterization of an aminopeptidase associated with larval molting.
AuthorsHong X, Bouvier J, Wong MM, Yamagata GY, McKerrow JH
JournalExp Parasitol
PubMed ID8454021
'The third-stage larvae (L3) of the filarid parasite Brugia pahangi were surveyed for protease activity beginning with release from the mosquito vector through molting to the fourth-stage larvae (L4). A metalloaminopeptidase with a substrate preference for phenylalanine was released during the molting process. A screen of aminopeptidase inhibitors identified H-boroPhenylalanine-(pinacol) ... More
Assay of coagulation proteases using peptide chromogenic and fluorogenic substrates.
AuthorsLottenberg R, Christensen U, Jackson CM, Coleman PL
JournalMethods Enzymol
PubMed ID6210826
Phylogenetic conservation of cysteine proteinases. Cloning and expression of a cDNA coding for human cathepsin S.
AuthorsWiederanders B, Brömme D, Kirschke H, von Figura K, Schmidt B, Peters C
JournalJ Biol Chem
PubMed ID1377692
'A 1.8-kilobase full-length cDNA of human cathepsin S, a lysosomal cysteine proteinase, has been isolated. The single long open reading frame encodes a polypeptide of 331 amino acids consisting of a 15-amino acid NH2-terminal signal peptide, a propeptide of 99 amino acids, and a mature polypeptide of 217 amino acids. ... More
Distinct properties of prohormone thiol protease (PTP) compared to cathepsins B, L, and H: evidence for PTP as a novel cysteine protease.
AuthorsAzaryan AV, Hook VY
JournalArch Biochem Biophys
PubMed ID7944391
'The prohormone thiol protease (PTP) has been demonstrated as a major processing enzyme involved in converting the enkephalin precursor to active opiate enkephalin peptides. In this report, PTP was distinguished from other mammalian cysteine proteases, cathepsins B, L, and H, with regard to selectivity for monobasic and paired basic residue-containing ... More
Human cathepsin O. Molecular cloning from a breast carcinoma, production of the active enzyme in Escherichia coli, and expression analysis in human tissues.
AuthorsVelasco G, Ferrando AA, Puente XS, Sánchez LM, López-Otín C
JournalJ Biol Chem
PubMed ID7929457
'A cDNA encoding a novel member of the cysteine proteinase family of proteins has been cloned from a human breast carcinoma cDNA library, by using a polymerase chain reaction-based cloning strategy. The isolated cDNA contains an open reading frame coding for a polypeptide of 321 amino acids that has been ... More
New fluorogenic peptide substrates for plasmin.
AuthorsKato H, Adachi N, Ohno Y, Iwanaga S, Takada K, Sakakibara S
JournalJ Biochem (Tokyo)
PubMed ID6447693
'Fluorogenic peptides, peptidyl-4-methylcoumaryl-7-amides (MCA), containing COOH-terminal lysine residues, were newly synthesized and tested as substrates for plasmin. Among six peptidyl-MCA''s, Boc-Val-Leu-Lys-MCA and Boc-Glu-Lys-Lys-MCA were found to be useful for the specific and sensitive assay of plasmin. The Km values estimated from Line-weaver-Burk plots for these substrates using human and bovine ... More
Adult Schistosoma mansoni express cathepsin L proteinase activity.
AuthorsSmith AM, Dalton JP, Clough KA, Kilbane CL, Harrop SA, Hole N, Brindley PJ
JournalMol Biochem Parasitol
PubMed ID7838171
'This report presents the deduced amino acid sequence of a novel cathepsin L proteinase from Schistosoma mansoni, and describes cathepsin L-like activity in extracts of adult schistosomes. Using consensus primers specific for cysteine proteinases, gene fragments were amplified from adult S. mansoni cDNA by PCR and cloned. One of these ... More
Importance of hydrogen-bonding interactions involving the side chain of Asp158 in the catalytic mechanism of papain.
AuthorsMénard R, Khouri HE, Plouffe C, Laflamme P, Dupras R, Vernet T, Tessier DC, Thomas DY, Storer AC
JournalBiochemistry
PubMed ID2036422
'In a previous study, it was shown that replacing Asp158 in papain by Asn had little effect on activity and that the negatively charged carboxylate of Asp158 does not significantly stabilize the active site thiolate-imidazolium ion pair of papain (Ménard et al., 1990). In this paper, we report the kinetic ... More
Membrane permeable fluorogenic rhodamine substrates for selective determination of cathepsin L.
AuthorsAssfalg-Machleidt I, Rothe G, Klingel S, Banati R, Mangel WF, Valet G, Machleidt W
JournalBiol Chem Hoppe Seyler
PubMed ID1515071
'The dipeptidyl rhodamine diamide substrates (Z-Phe-Arg)2-R110 and (Z-Arg-Arg)2-R110 are 820- and 360-fold more selective for cathepsin L than for cathepsin B allowing a sensitive determination of cathepsin L activity in the presence of high activity of cathepsin B. The results obtained with cell lysates suggest that the cysteine proteinase activity ... More
[Photometric method of determination of the amidase activity of proteinases using 4-methylcoumaryl-7-amide substrates]
AuthorsPozdnev VF, Rabinovich SE, Paskhina TS
JournalBioorg Khim
PubMed ID2400405
'It was shown that 7-amino-4-methylcoumarin (MC-amine), resulted from the enzymatic hydrolysis of 4-methylcoumaryl-7-amide (MC-amide) peptide substrates, may be estimated not only fluorometrically but also photometrically. A photometric method for estimating activity of tissue kallikrein (EC 3.4.21.35) and urokinase (EC 3.4.21.31) is suggested using Z-Phe-Arg-NHMC and Z-Gly-Gly-Arg-NHMC, respectively, as substrates. Kinetic ... More
The role of aspartic and cysteine proteinases in albumin degradation by rat kidney cortical lysosomes.
AuthorsBaricos WH, Zhou YW, Fuerst RS, Barrett AJ, Shah SV
JournalArch Biochem Biophys
PubMed ID3304168
'We have investigated the degradation of 125I-labeled bovine serum albumin by lysates of rat kidney cortical lysosomes. Maximal degradation of albumin occurred at pH 3.5-4.2, with approximately 70% of the maximal rate occurring at pH 5.0. Degradation was proportional to lysosomal protein concentration (range 100-600 micrograms) and time of incubation ... More
Cathepsin G.
AuthorsBarrett AJ
JournalMethods Enzymol
PubMed ID7341917
Cathepsin B, Cathepsin H, and cathepsin L.
AuthorsBarrett AJ, Kirschke H
JournalMethods Enzymol
PubMed ID7043200
Fluorimetric assays for cathepsin B and cathepsin H with methylcoumarylamide substrates.
AuthorsBarrett AJ
JournalBiochem J
PubMed ID6897924
Benzyloxycarbonyl-phenylalanyl-arginine 4-methyl-7-coumarylamide was found to be an excellent substrate for the fluorimetric assay of cathepsin B, and arginine 4-methyl-7-coumarylamide for cathepsin H. Procedures were developed that are very convenient, and avoid the hazards associated with the use of naphthylamides. ... More
Specific assay method for the activities of cathepsin L-type cysteine proteinases.
AuthorsInubushi T, Kakegawa H, Kishino Y, Katunuma N
JournalJ Biochem (Tokyo)
PubMed ID7822244
We have established a new differential assay method for cathepsin L-type proteinases using specific inhibitors, E-64 for all cysteine proteinases, CA-074 for cathepsin B, and PLCPI for cathepsin L-type proteinases with Z-Phe-Arg-MCA as the substrate. The value of cathepsin B calculated by this method did not coincide with value assayed ... More
New fluorogenic substrates for alpha-thrombin, factor Xa, kallikreins, and urokinase.
AuthorsMorita T, Kato H, Iwanaga S, Takada K, Kimura T
JournalJ Biochem (Tokyo)
PubMed ID591514
Twenty peptide-4-methylcoumarin amides (MCA) were newly synthesized and tested as possible substrates for alpha-thrombin, factor Xa, kallikreins, urokinase, and plasmin. These fluorogenic peptides contained arginine-MCA as the carboxyl-terminus. Release of 7-amino-4-methylcoumarin was determined fluorometrically. Of these peptides, the following were found to be specific substrates for individual enzymes: Boc-Val-Pro-Arg-MCA for ... More
Purification of a 29-kDa hemocyte proteinase of Sarcophaga peregrina.
AuthorsKurata S, Saito H, Natori S
JournalEur J Biochem
PubMed ID1541301
Previously, we suggested the participation of a hemocyte proteinase in the dissociation of fat body of Sarcophaga peregrina (flesh fly) at metamorphosis. We have now purified this proteinase to near homogeneity from pupal hemocytes. It is a cysteine proteinase with a molecular mass of 29 kDa and has a unique ... More
Purification of the complex of cathepsin L and the MHC class II-associated invariant chain fragment from human kidney.
AuthorsOgrinc T, Dolenc I, Ritonja A, Turk V
JournalFEBS Lett
PubMed ID8282126
The complex of cathepsin L and the fragment of the MHC class II-associated invariant chain was purified from human kidney. M(r) of the complex, as determined by gel filtration, is about 40,000. Both components were identified by amino acid and sequence analyses. The bound invariant chain fragment is almost identical ... More
Cathepsin B of Schistosoma mansoni. Purification and activation of the recombinant proenzyme secreted by Saccharomyces cerevisiae.
AuthorsLipps G, Füllkrug R, Beck E
JournalJ Biol Chem
PubMed ID8576174
Procathepsin B from the parasitic trematode Schistosoma mansoni was expressed as a glycosylation-minus mutant in yeast cells and purified by means of a histidine affinity tag which was added to the carboxyl terminus of the recombinant protein. The purified zymogen underwent autoprocessing but required an assisting protease for activation. Pepsin-activated ... More
A proteolytic activity enhanced by arsenite in Chinese hamster ovary cells: possible involvement in arsenite-induced cell killing.
AuthorsYih LH, Lee TC
JournalBiochem Biophys Res Commun
PubMed ID8048913
Treatment of Chinese hamster ovary (CHO-K1) cells with 10 microM sodium arsenite for 24 h resulted in enhancement of a proteolytic activity toward the chromogenic substrate CBZ-Phe-Arg-AMC. Presence of dithiothreitol and a pH between 4 and 6 were required for displaying its full hydrolytic activity. According to its substrate- and ... More
Potent inactivation of cathepsins S and L by peptidyl (acyloxy)methyl ketones.
AuthorsBrömme D, Smith RA, Coles PJ, Kirschke H, Storer AC, Krantz A
JournalBiol Chem Hoppe Seyler
PubMed ID8074807
Peptidyl (acyloxy)methyl ketones (Z-Aa-Aa-CH2-O-CO-R), a new class of irreversible inhibitors whose chemical reactivity can be modulated by varying the substitution pattern of the carboxylate leaving group, are shown to be extremely potent inactivators of the lysosomal cysteine proteinases cathepsin L and cathepsin S. The highest k2/Ki values measured were found ... More
Anti-peptide antibodies to cathepsins B, L and D and type IV collagenase. Specific recognition and inhibition of the enzymes.
AuthorsCoetzer TH, Elliott E, Fortgens PH, Pike RN, Dennison C
JournalJ Immunol Methods
PubMed ID1847962
Anti-peptide antibodies were raised against synthetic peptides selected from the sequences of human cathepsins B and L, porcine cathepsin D and human type IV collagenase. Sequences were selected from the active site clefts of the cathepsins in the expectation that these would elicit immunoinhibitory antibodies. In the case of type ... More
The specificity and elastinolytic activities of bovine cathepsins S and H.
AuthorsXin XQ, Gunesekera B, Mason RW
JournalArch Biochem Biophys
PubMed ID1444473
Cathepsins S and H were purified from bovine spleen and their catalytic properties compared. The enzymes were shown to be similar by chromatographic properties and by the ability to hydrolyze Bz-Phe-Val-Arg-NHMec. They could however be distinguished by the fact that cathepsin S reacted with Z-[125I]Tyr-Ala-CHN2 and hydrolyzed Z-Phe-Arg-NHMec whereas cathepsin ... More
Cathepsin L proteinase secreted by Fasciola hepatica in vitro prevents antibody-mediated eosinophil attachment to newly excysted juveniles.
AuthorsCarmona C, Dowd AJ, Smith AM, Dalton JP
JournalMol Biochem Parasitol
PubMed ID8114830
Cathepsin L-like activity was demonstrated in the excretory/secretory (E/S) products of Fasciola hepatica newly excysted juveniles (NEJ), 3-week-old, 5-week-old and mature flukes using the fluorogenic substituted 7-amino-4-methylcoumarin substrates Z-phe-arg-AMC, Z-arg-arg-AMC and Z-arg-AMC. Gelatin-substrate polyacrylamide gel analysis revealed that the E/S from each of these stages contained multiple proteolytic enzymes; however, ... More
Purification and characterization of cathepsin B from monkey skeletal muscle.
AuthorsHirao T, Hara K, Takahashi K
JournalJ Biochem (Tokyo)
PubMed ID6725239
Cathepsin B was purified about 11,000-fold from monkey skeletal muscle by ammonium sulfate fractionation and sequential column chromatographies monitored by assaying of Z-Phe-Arg-MCA hydrolase activity. The purified enzyme gave a single protein band on SDS-polyacrylamide gel electrophoresis, and its molecular weight was estimated to be 24,000 by gel filtration. It ... More
Putative N-terminal splitting enzyme of amyloid A4 peptides is the multicatalytic proteinase, ingensin, which is widely distributed in mammalian cells.
AuthorsIshiura S, Tsukahara T, Tabira T, Sugita H
JournalFEBS Lett
PubMed ID2573543
The main characteristic changes observed in Alzheimer's disease (AD) are the presence of neurofibrillary tangles and the deposition of amyloid A4 peptides. The most abundant amyloid A4 peptide species in AD (which we tentatively named A4') is composed of 39 amino acids, which is devoid of the 3 N-terminal amino ... More
Endopeptidase activity of cathepsin C, dipeptidyl aminopeptidase I, from bovine spleen.
AuthorsKuribayashi M, Yamada H, Ohmori T, Yanai M, Imoto T
JournalJ Biochem (Tokyo)
PubMed ID8514733
By employing various synthetic substrates, as well as soluble denatured protein substrate (TAP-lysozyme) and its derivatives, endopeptidase activity of cathepsin C, dipeptidyl aminopeptidase I [EC 3.4.14.1], from bovine spleen was investigated. Cathepsin C efficiently degraded Z-Phe-Arg-MCA, Pro-Phe-Arg-MCA, and Suc-Leu-Leu-Val-Tyr-MCA. This endopeptidase activity required sulfhydryl reagents and halide ions, as in ... More
Assessment of cathepsin L activity by use of the inhibitor CA-074 compared to cathepsin B activity in human lung tumor tissue.
AuthorsWerle B, Ebert W, Klein W, Spiess E
JournalBiol Chem Hoppe Seyler
PubMed ID7612192
In a series of pairs of lung tumor tissue and non-tumor lung parenchyma from 50 patients, the activity of cathepsin L was measured with Z-Phe-Arg-AMC using the inhibitor CA-074 to delimitate from cathepsin B activity also present in the tissue extracts. Cathepsin B was assessed in the same samples with ... More
Engineering nitrile hydratase activity into a cysteine protease by a single mutation.
AuthorsDufour E, Storer AC, Ménard R
JournalBiochemistry
PubMed ID8845364
A peptide nitrile hydratase activity has been engineered into the cysteine protease papain by a single carefully selected mutation at the active site of the enzyme. The papain variant Gln19Glu hydrolyzes the substrate MeOCO-PheAla-CN to the corresponding amide with a kcat/KM value of 1.15 x 10(3) M-1 s-1. The reaction ... More
Rat brain cathepsin L: characterization and differentiation from cathepsin B utilizing opioid peptides.
AuthorsMarks N, Berg MJ
JournalArch Biochem Biophys
PubMed ID3688881
The specificity of purified rat brain cathepsin L (EC 3.4.22.15) was mapped by the use of synthetic and opioid peptides and some properties were compared to rat brain cathepsin B, rat kidney cathepsin L, and bovine spleen cathepsin C. Brain and kidney cathepsin L cleaved leucine or methionine enkephalin (LE ... More
Lysine is a common determinant for mannose phosphorylation of lysosomal proteins.
AuthorsCuozzo JW, Sahagian GG
JournalJ Biol Chem
PubMed ID8182054
The phosphorylation of lysosomal enzymes on high mannose residues is the first step in the targeting of these enzymes to lysosomes in a wide range of mammalian cells. Phosphorylated lysosomal enzymes bind to mannose 6-phosphate receptors, which divert them from the secretory pathway and direct them toward the lysosome. We ... More
Evidence for the interaction of valine-10 in cystatin C with the S2 subsite of cathepsin B.
AuthorsLindahl P, Ripoll D, Abrahamson M, Mort JS, Storer AC
JournalBiochemistry
PubMed ID8155656
The interactions between wild-type or mutant recombinant forms of human cystatin C and rat cathepsin B were characterized by measuring progress curves for substrate hydrolysis in the presence of inhibitor. The investigation was guided by the use of computer modeling and explores the possibility that amino acid residues in the ... More
The role of cysteine proteases in hypoxia-induced rat renal proximal tubular injury.
AuthorsEdelstein CL, Wieder ED, Yaqoob MM, Gengaro PE, Burke TJ, Nemenoff RA, Schrier RW
JournalProc Natl Acad Sci U S A
PubMed ID7644473
The role of the lysosomal proteases cathepsins B and L and the calcium-dependent cytosolic protease calpain in hypoxia-induced renal proximal tubular injury was investigated. As compared to normoxic tubules, cathepsin B and L activity, evaluated by the specific fluorescent substrate benzyloxycarbonyl-L-phenylalanyl-L-arginine-7-amido-4-methylcoumarin, was not increased in hypoxic tubules or the medium ... More
L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) and its analogues as inhibitors of cysteine proteinases including cathepsins B, H and L.
AuthorsBarrett AJ, Kembhavi AA, Brown MA, Kirschke H, Knight CG, Tamai M, Hanada K
JournalBiochem J
PubMed ID7044372
1. L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) at a concentration of 0.5 mM had no effect on the serine proteinases plasma kallikrein and leucocyte elastase or the metalloproteinases thermolysin and clostridial collagenase. In contrast, 10 muM-E-64 rapidly inactivated the cysteine proteinases cathepsins B, H and L and papain (t0.5 = 0.1-17.3s). The streptococcal cysteine ... More
Measurement of elastase and cysteine proteinases in synovial fluid of patients with rheumatoid arthritis, sero-negative spondylarthropathies, and osteoarthritis.
AuthorsHuet G, Flipo RM, Richet C, Thiebaut C, Demeyer D, Balduyck M, Duquesnoy B, Degand P
JournalClin Chem
PubMed ID1526000
Synovial fluid samples were collected from 45 patients with rheumatoid arthritis, spondylarthropathy, or osteoarthritis, to study their content of elastase (EC 3.4.21.37) and of cysteine proteinases (EC 3.4.22.1, 3.4.22.15). We measured both elastase complexed with alpha 1-proteinase inhibitor and elastase activity toward the substrate L-pyroglutamyl-L-prolyl-L-valine-p-nitroanilide. Cysteine proteinase activities were measured ... More
Engineering the S2 subsite specificity of human cathepsin S to a cathepsin L- and cathepsin B-like specificity.
AuthorsBrömme D, Bonneau PR, Lachance P, Storer AC
JournalJ Biol Chem
PubMed ID7982933
The primary specificity of papain-like proteinases is largely determined by S2-P2 site interactions. According to the three-dimensional structure of a papain-inhibitor complex, the S2 subsite is defined by residues 67, 68, 133, 157, 160, and 205, with residues 133, 157, and 205 integrated into the wall and bottom of the ... More
The application of a novel biotinylated affinity label for the detection of a cathepsin B-like precursor produced by breast-tumour cells in culture.
AuthorsCullen BM, Halliday IM, Kay G, Nelson J, Walker B
JournalBiochem J
PubMed ID1575692
In this report we demonstrate how the recently developed biotinylated affinity label biotinyl-Phe-Ala-diazomethane (Bio-Phe-Ala-CHN2) [Cullen, McGinty, Walker, Nelson, Halliday, Bailie & Kay (1990) Biochem. Soc. Trans. 18, 315-316; Walker, Cullen, Kay, Halliday, McGinty & Nelson (1992) Biochem. J. 283, 449-453] can be used for the detection of a precursor form ... More
Secretion of cysteine proteinase activity by the zoonotic hookworm Ancylostoma caninum.
The zoonotic hookworm, Ancylostoma caninum, probably induces human eosinophilic enteritis by inducing allergic responses to its secretions. This species is already known to secrete metalloproteinases, but in other parasites, cysteine proteinases are involved in pathogenesis. We studied somatic extracts of A. caninum adults and infective larvae and adult excretory/secretory (ES) ... More
A limulus intracellular coagulation inhibitor type 2. Purification, characterization, cDNA cloning, and tissue localization.
AuthorsMiura Y, Kawabata S, Wakamiya Y, Nakamura T, Iwanaga S
JournalJ Biol Chem
PubMed ID7822280
We described in a foregoing report findings on serpin, a serine protease inhibitor, newly identified in horseshoe crab (Tachypleus tridentatus) hemocytes and we name it limulus intracellular coagulation inhibitor, LICI (Miura, Y., Kawabata, S., and Iwanaga, S. (1994) J. Biol. Chem. 269, 542-547). This serpin specifically inhibits limulus lipopolysaccharide-sensitive serine ... More
Purification and characterization of proteolytic enzymes of Leishmania mexicana mexicana amastigotes and promastigotes.
AuthorsPupkis MF, Coombs GH
JournalJ Gen Microbiol
PubMed ID6389769
Leishmania mexicana mexicana amastigote and promastigote soluble proteinases were purified using gel filtration and ion exchange chromatography. For the amastigotes, two main proteinase activity peaks were separated with both methods. These accounted for approximately 10% and 90% of the total activity. Characterization of the two activities for substrate specificity and ... More
Characterization of recombinant rat cathepsin B and nonglycosylated mutants expressed in yeast. New insights into the pH dependence of cathepsin B-catalyzed hydrolyses.
AuthorsHasnain S, Hirama T, Tam A, Mort JS
JournalJ Biol Chem
PubMed ID1537854
The cysteine proteinase rat cathepsin B was expressed in yeast in an active form and was found to be heterogeneously glycosylated at the consensus sequence for N-linked oligosaccharide substitution. Purified enzyme fractions containing the highest levels of glycosylation were shown to have reduced activity. A glycosylation minus mutant constructed by ... More