scholarly journals Evidence for polymorphonuclear-leucocyte-derived proteinases in arthritic cartilage

1981 ◽  
Vol 193 (1) ◽  
pp. 193-202 ◽  
Author(s):  
J D Sandy ◽  
A Sriratana ◽  
H L Brown ◽  
D A Lowther
Keyword(s):  
Guanidine Hydrochloride ◽  
Apparent Molecular Weight ◽  
Joint Inflammation ◽  
Polymorphonuclear Leucocyte ◽  
Soya Bean ◽  
Polymorphonuclear Leucocytes ◽  
Cartilage Proteoglycan ◽  
Marked Inhibition ◽  
Bean Trypsin Inhibitor ◽  
Proteoglycan Degradation

1. An enzyme that degrades proteoglycan at neutral pH was extracted with 4 M-guanidine hydrochloride from the articular cartilage of rabbits with antigen-induced arthritis. 2. The enzyme had an apparent molecular weight on Ultrogel AcA 54 of about 8000 and was optimally active at pH 7.5 in Tris/HCl buffer containing 0.2 M-NaCl. The partially purified preparation was totally inhibited by 0.01 mM-N-acetyldialanylprolylvalylchloromethane, severely inhibited by 2 mM-phenylmethanesulphonyl fluoride and soya-bean trypsin inhibitor (200 microgram/ml) and slightly inhibited by 10 mM-EDTA. Marked inhibition was also obtained with a cytosolic fraction prepared from rabbit polymorphonuclear leucocytes. 3. All properties of the enzyme were virtually identical with those of an ‘elastase-like’ proteinase that was isolated from rabbit polymorphonuclear-leucocyte granules. 4. The results are consistent with the idea that cartilage proteoglycan degradation in acute joint inflammation is due at least partly to the diffusion into the cartilage of proteinases derived from synovial-fluid polymorphonuclear leucocytes.

scholarly journals Proteoglycan-degrading enzymes of rabbit fibroblasts and granulocytes

1978 ◽  
Vol 173 (3) ◽  
pp. 949-958 ◽  
Author(s):  
Z Werb ◽  
J T Dingle ◽  
J J Reynolds ◽  
A J Barrett
Keyword(s):  
Trypsin Inhibitor ◽  
Gel Filtration ◽  
Synovial Fibroblasts ◽  
Enzymic Activity ◽  
Polymorphonuclear Leucocyte ◽  
Exchange Chromatography ◽  
Soya Bean ◽  
Polymorphonuclear Leucocytes ◽  
Neutral Proteinase ◽  
Bean Trypsin Inhibitor

A neutral proteinase secreted by rabbit synovial fibroblasts in parallel with specific collagenase was partially purified by ion-exchange chromatography. At pH 7.6 this proteinase degraded 35S-labelled bovine nasal proteoglycan and azo-casein. The enzymic activity was inhibited by EDTA, 1,10-phenanthroline and serum, whereas di-isopropyl phosphorofluoridate and soya-bean trypsin inhibitor had little effect. By gel filtration the apparent mol.wt. of the enzyme was 25000. The fibroblast neutral proteinase was compared with the proteoglycan-degrading neutral proteinases of rabbit polymorphonuclear-leucocyte granules. Two distinct activities were found in the granules: one was inhibited by soya-bean trypsin inhibitor and the other by EDTA. The proteoglycan-degrading proteinases of rabbit fibroblasts and polymorphonuclear leucocytes at acid pH also were examined. Both cathepsin D and a thiol-dependent proteinase contributed to the degradation of proteoglycan at pH 4.5.

Cold Promoted Activation of Factor VII VI. Effect of Inhibitors

1973 ◽  
Vol 29 (03) ◽  
pp. 633-643
Author(s):  
H Gjønnæss
Keyword(s):  
Soya Bean ◽  
Factor Vii ◽  
Plasma Kallikrein ◽  
Contact Activation ◽  
Activation Reaction ◽  
Hexadimethrine Bromide ◽  
Bean Trypsin Inhibitor ◽  
Plasma Arginine ◽  
Kallikrein Activity ◽  
Effect Of Inhibitors

SummaryThe cold promoted activation of factor VII occurs in parallel with an activation of a plasma arginine esterase, and, on inhibition of the cold activation of factor VII, the esterase activation also decreased. The inhibitor pattern supported our theory that the arginine esterase that is activated in the cold activation of factor VII is plasma kallikrein.The cold activation of factor VII was completely inhibited with soya bean trypsin inhibitor in doses that did not interfere with the contact activation. On the other hand, inhibition of the contact activation with hexadimethrine bromide did not interfere with the cold activation of factor VII except when this was kaolin induced. Contact and cold activation therefore appear to represent two different pathways for the activation of factor VII. The cold activation reaction is probably mediated by the activation of plasma prekallikrein, and inhibition of the plasma kallikrein activity correlates with the inhibition of the cold promoted activation of factor VII.

Activation of latent collagenase from polymorphonuclear leukocytes by cathepsin G

1979 ◽  
Vol 44 (10) ◽  
pp. 3177-3182 ◽  
Author(s):  
Mária Stančíková ◽  
Karel Trnavský
Keyword(s):  
Affinity Chromatography ◽  
Trypsin Inhibitor ◽  
Polymorphonuclear Leukocytes ◽  
Soya Bean ◽  
Cathepsin G ◽  
Sepharose Column ◽  
Bean Trypsin Inhibitor ◽  
Human Polymorphonuclear Leukocytes

Cathepsin G was isolated from human polymorphonuclear leukocytes and purified by affinity chromatography on Antilysin-Sepharose column. Purified enzyme activated later collagenase isolated from leukocytes. Activation at 36°C was maximal after 30 min incubation. Inhibitors of cathepsin G - soya-bean trypsin inhibitor, diisopropyl phosphofluoridate and Antilysin were active in inhibiting the activation of latent collagenase by cathepsin G.

scholarly journals The immunologic detection and characterization of cartilage proteoglycan degradation products in synovial fluids of patients with arthritis

1987 ◽  
Vol 30 (5) ◽  
pp. 519-529 ◽  
Author(s):  
James Witter ◽  
Peter J. Roughley ◽  
Carolyn Webber ◽  
Nancy Roberts ◽  
Edward Keystone ◽  
...  
Keyword(s):  
Degradation Products ◽  
Cartilage Proteoglycan ◽  
Synovial Fluids ◽  
Proteoglycan Degradation

scholarly journals Lysosomal cysteine endopeptidases mediate interleukin 1-stimulated cartilage proteoglycan degradation

1992 ◽  
Vol 287 (2) ◽  
pp. 657-661 ◽  
Author(s):  
D J Buttle ◽  
J Saklatvala
Keyword(s):  
Interleukin 1 ◽  
Active Form ◽  
Growth Factor Receptor ◽  
Test System ◽  
Significant Inhibition ◽  
Cartilage Explants ◽  
Cartilage Proteoglycan ◽  
Proteoglycan Degradation ◽  
Cysteine Endopeptidases ◽  
Proteoglycan Release

The peptidyl diazomethane inactivator of cysteine endopeptidases, benzyloxycarbonyl-Tyr-Ala-CHN2, was tested as an inhibitor of interleukin 1 alpha-stimulated release of proteoglycan from bovine nasal septum cartilage explants. Like the previously tested epoxidyl peptide proinhibitor trans-epoxysuccinyl-leucylamido-(3-methyl)butane ethyl ester, it proved to be an effective inhibitor of proteoglycan release from cartilage, with significant inhibition at a concentration of 1 microM. The inhibition did not seem to be due to a general toxic effect. The rates of inactivation of the bovine cysteine endopeptidases by the peptidyl diazomethane, the epoxidyl peptide proinhibitor and its active form were determined. Benzyloxycarbonyl-Tyr-Ala-CHN2 proved to be a rapid inactivator of cathepsins L, S and B, but reacted much more slowly with cathepsin H and calpain. Thus it would appear that the latter two enzymes are not implicated in proteoglycan release in our test system. The peptidyl diazomethane and epoxidyl peptide proinhibitor (above) were also tested for their effects on three other interleukin 1-mediated cellular events, namely epidermal growth factor receptor transmodulation, and interleukin 6 and prostaglandin E2 production. In all cases the inactivators did not interfere with the response to interleukin 1 in human gingival fibroblasts. We conclude that one or more of the lysosomal cysteine endopeptidases cathepsins B, L and S mediate interleukin 1-stimulated cartilage proteoglycan degradation without affecting signal transduction.

scholarly journals Cytokine-induced cartilage proteoglycan degradation is mediated by aggrecanase

1998 ◽  
Vol 6 (3) ◽  
pp. 214-228 ◽  
Author(s):  
Elizabeth C. Arner ◽  
Clare E. Hughes ◽  
Carl P. Decicco ◽  
Bruce Caterson ◽  
Micky D. Tortorella
Keyword(s):  
Cartilage Proteoglycan ◽  
Proteoglycan Degradation

Physikalische, chemische und biologische Charakterisierung von menschlichem Choriongonadotropin

1968 ◽  
Vol 23 (11) ◽  
pp. 1412-1426 ◽  
Author(s):  
H. D. Schlumberger
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Biological Activities ◽  
Guanidine Hydrochloride ◽  
Group Analysis ◽  
Residual Activity ◽  
Apparent Molecular Weight ◽  
Terminal Amino Acid ◽  
Original Activity ◽  
Terminal Amino

Purification of commercially available HCG preparations with DEAE Sephadex A 50 and Sephadex G 100 column chromatography gave homogeneous fractions having specific biological activities four fold those of the starting materials. The purified HCG has ICSH characteristics and, in high doses, a definite FSH effect is present.Chemical analysis of HCG showed it to contain 29% carbohydrates and 69% peptides. The C-terminal amino acid of the peptide chain was found to be serine, but the N-terminal amino acid could not be determined with normal methods. A molecular weight of 22000 — 27000 daltons was obtained by quantitative end group analysis. Ultracentrifugation experiments in 4 m guanidine hydrochloride gave a molecular weight of 27200 daltons, but in neutral saline solutions at HCG concentrations above 2 mg/ml the apparent molecular weight was higher and indicated dimer formation. A dissociation constant of 10-5 mol/l was estimated for the monomer-dimer equilibrium. Since biological activity is found with 0.1 to 0.5 µg, it was concluded that the HCG monomer is the active entity.The purified HCG is stable from pH 4.5 to pH 10 for 6 hours at 37 °C. At pH 2.5 only 5 to 10% of the original activity is retained. HCG is rapidly inactivated at 100 °C, but a residual activity of 6 — 10% remained after 30 minutes at 80 °C. No activity was lost after 30 minutes incubation at 60 °C.

Evidence for a membrane-bound form of a bacteriocin of Bacteroides uniformis Tl-1

1984 ◽  
Vol 30 (2) ◽  
pp. 268-272 ◽  
Author(s):  
Sandra L. Austin-Prather ◽  
S. James Booth
Keyword(s):  
Molecular Weight ◽  
Outer Surface ◽  
Guanidine Hydrochloride ◽  
Membrane Vesicles ◽  
Apparent Molecular Weight ◽  
Whole Cells ◽  
Membrane Bound ◽  
Bleb Formation

The bacteriocin of Bacteroides uniformis Tl-1 had previously been reported to have a molecular weight of ≥ 300 000. Reexamination of the B. uniformis bacteriocin revealed that the bacteriocin was found in association with membrane vesicles which had been released by bleb formation from the outer surface of the B. uniformis cells. The bacteriocin could be released from whole cells or purified membrane vesicles by treatment with 6 M guanidine hydrochloride or 7 M urea and had an apparent molecular weight of 5000–6200.

scholarly journals The recovery of human polymorphonuclear leucocytes from sublytic complement attack is mediated by changes in intracellular free calcium

1985 ◽  
Vol 231 (1) ◽  
pp. 205-208 ◽  
Author(s):  
B P Morgan ◽  
A K Campbell
Keyword(s):  
Calcium Ion ◽  
Membrane Attack Complex ◽  
Polymorphonuclear Leucocyte ◽  
Intracellular Free Calcium ◽  
Ion Concentration ◽  
Free Calcium ◽  
Polymorphonuclear Leucocytes ◽  
Recovery Processes ◽  
Rapid Removal ◽  
Calcium Ion Concentration

Using polymorphonuclear leucocyte-erythrocyte ghost hybrids entrapping the calcium-activated photoprotein obelin, we have demonstrated that sublytic amounts of the complement membrane attack complex induce a rapid but transient increase in intracellular free calcium ion concentration ([Ca2+]i). This increase in [Ca2+]i occurs prior to, and is required for, rapid removal of membrane attack complexes from the cell surface. The increase in [Ca2+]i is not only due to increased influx from outside the cell, but also results from mobilization of intracellular stores. The possible mechanism of mobilization of calcium, and the importance of an increase in [Ca2+]i as a mediator of recovery processes in nucleated cells, are discussed.

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