Biosynthesis and secretion of procollagenase by rabbit synovial fibroblasts. Inhibition of procollagenase secretion by monensin and evidence for glycosylation of procollagenase

Nagase, Hideaki; Brinckerhoff, Constance E.; Vater, Carol A.; Harris, Edward D.

doi:10.1042/bj2140281

Cited by 93 publications

(33 citation statements)

References 45 publications

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“…of an amino-terminal propeptide (maintaining latency), a catalytic domain harbouring the zinc-and calciumsites, and a C-terminal hemopexin like domain, which is important for the unique ability of FIB-CL and PMNL-CL to cleave collagen [3-71. In contrast to the FIB pro-CL, which is secreted immediately after synthesis [8], PMNL pro-CL is stored as a highly glycosylated protein within specific granules of neutrophils and can be secreted as an inactive precursor after initiation by inflammatory mediators [9-l 11. Conversion of the latent proenzyme to the active enzyme is a crucial step in the neutrophil-mediated initiation of collagenolysis during inflammatory connective tissue turnover.…”

Section: Introductionmentioning

confidence: 99%

Structural implications for the role of the N terminus in the ‘superactivation’ of collagenases

et al. 1994

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For the collagenases PMNL-CL and FIB-CL, the presence of the N-terminal Phe79 correlates with an increase in proteolytic activity. We have determined the X-ray crystal structure of the recombinant Phe79-Gly"2 catalytic domain of human neutrophil collagenase (PMNL-CL, MMP-8) usin the recently solved model of the Met80_Gly242 form for phasing and subsequently refined it to a final crystalographic R-factor of 18.0% at 2.5 x resolution. The PMNL-CL catalytic domain is a spherical molecule with a flat active site cleft separating a smaller C-terminal subdomain from a bigger N-terminal domain, that harbours two zinc ions, namely a 'structural' and a 'catalytic' zinc, and two calcium ions. The N-terminal segment prior to Pros6, which is disordered in the Met*'-Gly 242 form packs against a concave hydrophobic surface made by the C-terminal helix. The , N-terminal Phe79 ammonium group makes a salt link with the side chain carboxylate group of the strictly conserved AsP'~~. Stabilization of the catalytic site might be conferred via strong hydrogen bonds made by the adjacent, likewise strictly conserved Asp233 with the characteristic 'Met-turn', which forms the base of the active site residues.

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Section: Introductionmentioning

confidence: 99%

Structural implications for the role of the N terminus in the ‘superactivation’ of collagenases

et al. 1994

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“…Their primary structures have been elucidated by analysis of their respective cDNAs, demonstrating a close structural relationship between these homologous enzymes Hasty et al, 1990). The fibroblast proenzyme is secreted immediately after synthesis in vitro and in vivo (Nagase et al, 1983), and only a minor amount of the proenzyme is posttranslationally processed by glycosylation . In contrast, the neutrophil procollagenase is stored as a glycosylated protein within the specific granules ofthe neutrophils (Murphy et al, 1977;Knauper et al, 1990a).…”

Section: Introductionmentioning

confidence: 99%

Direct activation of human neutrophil procollagenase by recombinant stromelysin

Knäuper

Wilhelm

Seperack

et al. 1993

Biochemical Journal

149

105

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Human neutrophil procollagenase was activated by incubation with recombinant active stromelysin. Activation was achieved by cleavage of the Gly78-Phe79 peptide bond at the end of the propeptide domain in a single-step activation mechanism. In addition, accelerated activation was achieved when N-terminally truncated, latent collagenase (with Phe49 as its N-terminal residue) was incubated with recombinant active stromelysin. Determination of the specific activity of recombinant-stromelysin-activated neutrophil collagenase with dinitrophenyl-octapeptide or type I collagen demonstrated the generation of high specific activity. The specific activity of stromelysin-activated enzyme was considerably higher than that of trypsin- or HgCl2-activated collagenase. Thus human neutrophil collagenase is superactivated, like the homologous fibroblast collagenase [Murphy, Cockett, Stephens, Smith and Docherty (1987) Biochem. J. 248, 265-268]. The occurrence of Phe79 at the N-terminus of the neutrophil collagenase seemed to be critical for superactivation, which is in agreement with data published by Suzuki, Enghild, Morodomi, Salvesen and Nagase [(1990) Biochemistry 29, 10261-10270] on fibroblast collagenase.

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“…Upon addition of a stimulus to cultures of rabbit synovial fibroblasts, increased collagenase mRNA is detected in the cell by 5 hr and collagenase protein appears in the culture medium by 10 hr (14,17,19). Studies on the biosynthesis of both synovial and skin collagenase show that the time required for synthesis and secretion is <1 hr (13,16). Synovial cell collagenase is secreted predominantly as latent procollagenase of Mr 57,000, along with a small amount of a glycosylated species of Mr 61,000 (16,20).…”

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confidence: 99%

“…In contrast, fibroblast (e.g., skin and synovial cell) collagenase is not stored. Addition of an inducer stimulates transcription of collagenase mRNA followed by rapid synthesis and secretion of the enzyme (13)(14)(15)(16)(17).…”

mentioning

confidence: 99%

“…Studies on the biosynthesis of both synovial and skin collagenase show that the time required for synthesis and secretion is <1 hr (13,16). Synovial cell collagenase is secreted predominantly as latent procollagenase of Mr 57,000, along with a small amount of a glycosylated species of Mr 61,000 (16,20). Activation of the latent enzyme occurs through limited proteolysis (7) or through a specific activator protein (7,21) that is produced by skin, uterine, and synovial fibroblasts.…”

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confidence: 99%

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Autoregulation of collagenase production by a protein synthesized and secreted by synovial fibroblasts: cellular mechanism for control of collagen degradation.

Brinckerhoff¹,

Benoit²,

Culp³

1985

Proc. Natl. Acad. Sci. U.S.A.

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Conditioned medium taken from cultures of resting rabbit synovial fibroblasts contained a protein that prevented the synthesis of the neutral proteinase collagenase. Conditioned medium was concentrated 10-fold and placed on cultures of rabbit synovial fibroblasts along with an inducer of collagenase (phorbol myristate acetate or latex particles) and[3H]leucine. Collagenase production was measured by immunoprecipitation of culture medium with monospecific antibody. Gel filtration showed that the inhibitory factor had Mrs of 12,500, 25,000-50,000, and 150,000, suggesting that the protein may exist as aggregates. Activity was destroyed by boiling, by trypsin, and by dithiothreitol. Production of the inhibitory protein was prevented by cycloheximide. Isoelectric focusing purified the protein 100-to 150-fold and revealed pls in the range of 3.2-3.7. Glycosylation was demonstrated by binding to Con A-Sepharose. Our data indicate that rabbit synovial fibroblasts autoregulate collagenase production and suggest that the low levels of collagenase seen in resting cultures result from an active suppression of collagenase synthesis.The interstitial collagens are the body's most abundant structural proteins, and collagen remodeling occurs in a number of normal processes such as uterine resorption and wound healing (1-3) and in diseases such as rheumatoid arthritis and tumor invasion (4, 5) (for reviews, see refs. 6 and 7). The fact that collagen breakdown can be initiated only by the metalloproteinase collagenase (EC 3.4.24.7) makes this enzyme rate-limiting in collagenolysis and assures the importance of cellular mechanisms regulating its production. Although collagenase is produced by a variety of mammalian cell types (6, 7), the mechanisms governing collagenase synthesis and secretion differ. In polymorphonuclear leukocytes (8, 9) and monocytes/macrophages (10-12), collagenase is synthesized and stored within the cell in granules; secretion is regulated by factors controlling granule release rather than by those controlling enzyme synthesis (7). In contrast, fibroblast (e.g., skin and synovial cell) collagenase is not stored. Addition of an inducer stimulates transcription of collagenase mRNA followed by rapid synthesis and secretion of the enzyme (13-17).An abundant source of enzyme is that produced by fibroblasts derived from the synovial tissue lining the joints (4, 6, 7). Normally, these cells are quiescent, but proliferating synovial tissue in rheumatoid arthritis produces large quantities of collagenase, and the role of this enzyme in the destruction of connective tissues is well documented (4, 6, 7). The fact that these high levels of collagenase can be induced by treating cultures of resting synovial fibroblasts with a variety of stimuli makes these cells an attractive experimental model for studies on mechanisms controlling collagenase synthesis (14,(17)(18)(19).Over the past few years, considerable information has been obtained on the biosynthesis of synovial cell collagenase. Upon addition of a stimulus ...

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Biosynthesis and secretion of procollagenase by rabbit synovial fibroblasts. Inhibition of procollagenase secretion by monensin and evidence for glycosylation of procollagenase

Cited by 93 publications

References 45 publications

Structural implications for the role of the N terminus in the ‘superactivation’ of collagenases

Structural implications for the role of the N terminus in the ‘superactivation’ of collagenases

Direct activation of human neutrophil procollagenase by recombinant stromelysin

Autoregulation of collagenase production by a protein synthesized and secreted by synovial fibroblasts: cellular mechanism for control of collagen degradation.

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