Human leukocyte collagenase: Characterization of enzyme kinetics by a new method

Turto, Heikki; Lindy, Seppo; Uitto, Veli‐Jukka; Wegelius, Otto; Uitto, Jouni

doi:10.1016/0003-2697(77)90059-8

Cited by 68 publications

(37 citation statements)

References 34 publications

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“…First, the specificity of the collagenase was assessed by analyzing the degradative products of 3H-collagen on SDS-polyacrylamide slab gel electrophoresis. The pattern of 3H-labeled peptides showed the presence of distinct bands corresponding to al(1)" and d(I)* as well as ~l ( 1 )~ and c~2(1)~ peptides of type I collagen; these bands represent 3/4 and Y4 fragments of collagen a-chains, respectively (20) (Figure 4). Thus, the results indicate that the monocyte collagenase cleaves type I collagen in the same region as do other vertebrate type collagenases (21).…”

Section: Resultsmentioning

confidence: 99%

The production of collagenase by adherent mononuclear cells cultured from human peripheral blood

Louie

Weiss

Ryhänen

et al. 1984

Arthritis & Rheumatism

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Mononuclear cells were isolated from human peripheral blood by Ficoll-Hypaque centrifugation, and the cells adherent to plastic substrata were cultured in serum-free media supplemented with lactalbumin hydrolysate. These cell cultures, which consisted predominantly of monocyte-macrophages as judged by nonspecific esterase staining, accumulated collagenase in the medium. This collagenase resembled other vertebrate collagenases in that it cleaved native triple-helical type I collagen at a locus Y4-length away from the aminoterminal end of the molecule. The collagenase activity was inhibited by Na2EDTA, dithiothreitol, and fetal calf serum, while the addition of Ca++ or N-ethylmaleimide enhanced the enzyme activity. 1984. collagenase in the culture media was markedly enhanced by the incubation of cells with concanavalin A or phorbol myristic acetate. In the presence of cycloheximide, the levels of collagenase activity were markedly reduced, suggesting that active protein synthesis was required to express the enzyme activity. In additional experiments, monocytes were further purified by counterflow centrifugation-elutriation. The collagenase production was markedly increased in cultures enriched in monocyte-macrophages and devoid of polymorphonuclear leukocytes. The accumulation of collagenase in monocyte cultures incubated for 48 hours in the presence of concanavalin A or phorbol myristic acetate was of the same order of magnitude as in parallel cultures containing the same number of polymorphonuclear leukocytes purified by Ficoll-Hypaque centrifugation and Plasmagel sedimentation. The demonstration of collagenase activity in the monocyte cultures appears to reflect the increased diversity of monocyte functions which may play an important role in the tissue damage in chronic inflammatory diseases such as rheumatoid arthritis.In recent years, the monocyte-macrophage, the hallmark of chronic inflammatory cell infiltrates, has been found to possess an increased repertoire of secretory functions ( 1,2). Macrophages recovered from peritoneal exudate (3,4), pulmonary alveolar fluid (5,6), and tumor cell lines (7) have been found to secrete collagenase, as well as other proteolytic enzymes thought to mediate the degradation of connective tissue matrices (8). In this study, we have investigated collagenase production by monocyte cultures established from human peripheral blood. Because monocytes are precursors of tissue macrophages within rheumatoid and other chronic inflammatory lesions,

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Section: Resultsmentioning

confidence: 99%

The production of collagenase by adherent mononuclear cells cultured from human peripheral blood

Louie

Weiss

Ryhänen

et al. 1984

Arthritis & Rheumatism

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“…The enzyme reaction was terminated by boiling for 5 minutes after addition of sample buffer without 2-mercaptoethanol. Intact collagen a chains and their 3/4 (aA) and ?h (aB) degradation products were separated by 11% SDS-PAGE (41). The percentage of collagen degraded was estimated by image analysis (Molecular Analyst; Bio-Rad) of Coomassie blue-stained gels.…”

Section: Methodsmentioning

confidence: 99%

Matrix metalloproteinase 13 (collagenase 3) in human rheumatoid synovium

Lindy

Konttinen

Sorsa

et al. 1997

Arthritis & Rheumatism

196

115

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Objective. To show the eventual presence and extent of production of matrix metalloproteinase 13 (MMP-13, or collagenase 3) in rheumatoid synovial tissue samples and extracts, and to assess the inhibition characteristics of recombinant MMP-13.Methods. Immunohistochemical avidin-biotinperoxidase complex staininglmorphometry was used to analyze MMP-13-positive cells in situ. Neutral salt extraction of synovial tissue, electrophoresis of the extract in different buffer systems, and Western blotting were also used. The inhibitory properties of doxycycline, clodronate, pamidronate, and D-penicillamine for recombinant enzyme were determined with a soluble type I1 collagen assay.Results. MMP-13 was detected in fibroblast-and macrophage-like mononuclear cells in the synovial lining and stroma and in vascular endothelial cells. The overall expression of MMP-13 in these cells in the synovial stroma was high in rheumatoid arthritis (86 2 12%) compared with osteoarthritis (17 f 5%) patient samples (P = 0.0027). In a high-pH native electrophoresis gel, immunoreactivity to anti-MMP-1 and anti-MMP-13 were clearly separated, with anti-MMP-13-immunoreactive material migrating faster than anti-MMP-l-immunoreactive material. Finally, in contrast to MMP-1 and

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“…Saliva samples were assayed for interstitial collagenase activities by the quantitative SDS PAGE Laser densitometric modification [17][18][19]21,22] of the original assay developed by Turto et al [26]. Salivary samples were assayed for interstitial collagenase activities with (total collagenase activity) and without (endogenously active interstitial collagenase) 1 mM aminophenylmercuric acetate (APMA) treatment, an optimal organomercurial activator of both latent fibroblasl type and neutrophil interstitial collagenases.…”

Section: Sds-pagementioning

confidence: 99%

In vivo inhibition of human neutrophil collagenase (MMP-8) activity during long-term combination therapy of doxycycline and non-steroidal anti-inflammatory drugs (NSAID) in acute reactive arthritis

Lauhio

Salo

Ding

et al. 2008

Clinical &Amp; Experimental Immunology

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SUMMARYWe studied the in vivo effect of long-term doxycycline treatment combined with NSAID on human interstitial collagenases, other matrix melalloproteinases. serine proteinases, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and iactoferrin from saliva and serum during the course of acute reactive arthritis (ReA). Collagenase activity and serine proteases (elastase-like, cathepsin G-like and trypsin-like activities) of saliva {n = 10) and gelatinase, lacloferrin and TIMP-1 of saliva (/i =10) and serum (n -10) samples before and after 2 months doxycycline treatment, combined with NSAID, were studied by quantitative SDS-PAGE assay, ELISA assay and by spectrophotometric assay. The cellular source and molecular forms of salivary collagenase were characterized by immunoblotting using specific antisera. We found that activities of total and endogenousty active interstitial collagenase reduced significantly. The salivary collagenase was found to originate from neutrophils. No fragmentation of eilher pro 75-kD and active 65-kD MMP-8 was detected after 2 months doxycycline treatment. However, during 2 months doxycycline and NSAID treatment no reduction of salivary and serum gelatinase, Iactoferrin and TIMP-1-levels and salivary serine protease activities were delected. The in vivo inhibition ofcollagenase (MMP-8) activity during long-term doxycycline therapy in human saliva containing inflammatory exudate of ReA patients may contribute to the reduced tissue destruction observed in recent clinical and animal model studies in arthritides during long-term doxycycline/tetracycline treatment.Keywords doxycycline coliagenase/matrix metalloproteinase reactive arthritis collagenase/matrix metalloproteinase inhibition inflammatory diseases INTRODUCTION

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Human leukocyte collagenase: Characterization of enzyme kinetics by a new method

Cited by 68 publications

References 34 publications

The production of collagenase by adherent mononuclear cells cultured from human peripheral blood

The production of collagenase by adherent mononuclear cells cultured from human peripheral blood

Matrix metalloproteinase 13 (collagenase 3) in human rheumatoid synovium

In vivo inhibition of human neutrophil collagenase (MMP-8) activity during long-term combination therapy of doxycycline and non-steroidal anti-inflammatory drugs (NSAID) in acute reactive arthritis

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