Assessment of procollagen processing defects by fibroblasts cultured in the presence of dextran sulphate

Bateman, John F.; Golub, Suzanne B.

doi:10.1042/bj2670573

Cited by 51 publications

(25 citation statements)

References 34 publications

(46 reference statements)

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“…In this in vitro study, we evaluated the release of several cytokines and biological mediators from Caco-2 cells. In particular, DSS influences in vitro fibrinogenic-fibrolytic systems such as ß-FGF (21), collagen (22), procollagen (23), and bradykinin (24), or in vivo systems such as MMPs (25). As a result, DSS weakly induces the release of IL-8, IL-6 and TGF-ß1, and does not influence IL-10, MMP-1, u-PA, or bradykinin.…”

Section: Discussionmentioning

confidence: 99%

In vitro effects of dextran sulfate sodium on a Caco-2 cell line and plausible mechanisms for dextran sulfate sodium-induced colitis

Araki¹,

Sugihara²,

Hattori³

2006

Oncol Rep

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Abstract. Pathogenic mechanisms responsible for inflammatory bowel disease (IBD) are poorly understood. In an IBD animal model, the oral administration of polysaccharides such as dextran sulfate sodium (DSS) induces colitis, which exhibit several clinical and histological features for IBD. However, pathogenic factors in the development of colitis remain unclear. Therefore, we investigated possible mechanisms for DSS-induced colitis, and mainly focused on biological responses from an intestinal epithelial cell line, Caco-2. Cytotoxicity and cytokine release were measured using MTS assays and ELISA, respectively. The effect of DSS on the transepithelial electrical resistance (TEER) of Caco-2 cell monolayers was also evaluated. Cell cycle progression was estimated using antibodies directed against p53 and cdc-2 proteins. The generation of reactive oxygen species (ROS) was measured using a DCFH-DA method. Pyridylamino-DSS (PA-DSS) was used as a fluorometric label in order to investigate fluorescence-microscopically the location of DSS in Caco-2 cells. DSS induced cytotoxicity on Caco-2 cells at 5%. DSS also induced strong TEER decrease at 3%. DSS induced the weak release of IL-8, IL-6, and TGF-ß1. Remarkably DSS arrested Caco-2 cell cycle and reduced the intracellular generation of ROS. Under fluorescence microscopy, PA-DSS entered cells and bound to the nucleus, indicating this binding of DSS may be involved in the cell cycle arrest of Caco-2 cells. The cell cycle arrest and reduced intracellular generation of ROS may be involved during initiation or throughout the early stages of DSS-induced colitis.

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Section: Discussionmentioning

confidence: 99%

In vitro effects of dextran sulfate sodium on a Caco-2 cell line and plausible mechanisms for dextran sulfate sodium-induced colitis

Araki¹,

Sugihara²,

Hattori³

2006

Oncol Rep

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“…When produced by 293 cells grown in culture medium supplemented with fetal calf serum alone, recombinant ADAMTS-14 did not display any activity. However, upon limited digestion of the latent enzyme with trypsin or when recombinant ADAMTS-14 was extracted from cells cultivated in conditions known to promote metalloprotease activation (29,30), significant aminoprocollagen peptidase activity was evidenced. These results were further confirmed in a co-culture model.…”

Section: Figmentioning

confidence: 99%

Cloning and Characterization of ADAMTS-14, a Novel ADAMTS Displaying High Homology with ADAMTS-2 and ADAMTS-3

Colige¹,

Vandenberghe²,

Thiry³

et al. 2002

Journal of Biological Chemistry

184

129

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The processing of amino-and carboxyl-propeptides of fibrillar collagens is required to generate collagen monomers that correctly assemble into fibrils. Mutations in the ADAMTS2 gene, the aminopropeptidase of procollagen I and II, result in the accumulation of nonfully processed type I procollagen, causing human Ehlers-Danlos syndrome type VIIC and animal dermatosparaxis. In this study, we show that the aminopropeptide of type I procollagen can be cleaved in vivo in absence of ADAMTS-2 activity and that this processing is performed at the cleavage site for ADAMTS-2. In an attempt to identify the enzyme responsible for this alternative aminoprocollagen peptidase activity, we have cloned the cDNA and determined the primary structure of human and mouse ADAMTS-14, a novel ADAMTS displaying striking homologies with ADAMTS-2 and -3. The structure of the human gene, which maps to 10q21.3, and the mechanisms of generation of the various transcripts are described. The existence of two sites of initiation of transcription, in two different promoter contexts, suggests that transcripts resulting from these two sites can be differently regulated. The tissue distribution of AD-AMTS-14, the regulation of the gene expression by various cytokines and the activity of the recombinant enzyme are evaluated. The potential function of ADAMTS-14 as a physiological aminoprocollagen peptidase in vivo is discussed. ADAMTS1 (A Disintegrin and metalloprotease with thrombospondin type I repeats) is a novel family of metalloproteases found in vertebrates and invertebrates. These enzymes are related to ADAMs as judged from sequence homology and conserved domains such as a characteristic metalloprotease domain and a disintegrin-like module. However, they differ from ADAMs by their domain organization and the presence of distinct features. The most specific hallmark is the presence of a central thrombospondin type I repeat (TSPI) between the disintegrin-like module and the Cys-rich domain. All ADAMTS, except ADAMTS-4, contain also TSPI-like domains in varying numbers at the COOH terminus (1, 2). Currently, 12 ADAMTS from vertebrates and a few from invertebrates (Drosophila and Caenorhabditis elegans) have been described (1, 2). AD-AMTS-1, -4, and -5 are able to cleave proteoglycans and are probably involved in cartilage degradation during arthritis (3-5). ADAMTS-1 and -8 are potent anti-angiogenic molecules (6). Adamts1Ϫ/Ϫ mice display abnormal growth, defective fertility, and altered organ morphology and function (7). A C. elegans Adamts, gon-1, was found essential for gonadal morphogenesis (8). Both the metalloprotease domain and some TSPI-like repeats are required for the control of this process.The primarily described activity of ADAMTS-2 is to excise the aminopropeptide of type I and type II procollagens, explaining its former trivial name aminoprocollagen I/II peptidase (9, 10). Removal of the N-and C-propeptide of type I procollagen is required to generate collagen monomers able to assemble into elongated and cylindrical collagen fibers. H...

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“…Previous studies have shown that procollagen I processing in dermatosparactic fibroblasts is enhanced by including dextran sulfate (37,38) or polyethylene glycol (38) in the culture medium. However, despite the increased efficiency, processing was still incomplete (37,38).…”

Section: Adamts-2 and Adamts-3 Comprise Of A Structurally And Functiomentioning

confidence: 99%

Procollagen II Amino Propeptide Processing by ADAMTS-3

Fernandes¹,

Hirohata²,

Engle³

et al. 2001

Journal of Biological Chemistry

222

161

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The amino and carboxyl propeptides of procollagens I and II are removed by specific enzymes as a prerequisite for fibril assembly. Null mutations in procollagen I Npropeptidase (ADAMTS-2) cause dermatosparaxis in cattle and the Ehlers-Danlos syndrome (dermatosparactic type) in humans by preventing proteolytic excision of the N-propeptide of procollagen I. We have found that procollagen II is processed normally in dermatosparactic nasal cartilage, suggesting the existence of another N-propeptidase(s). We investigated such a role for ADAMTS-3 in Swarm rat chondrosarcoma RCS-LTC cells, which fail to process the procollagen II N-propeptide. Stable transfection of RCS-LTC cells with bovine ADAMTS-2 or human ADAMTS-3 partially rescued the processing defect, suggesting that ADAMTS-3 has procollagen II N-propeptidase activity. Human skin and skin fibroblasts showed 30-fold higher mRNA levels of ADAMTS-2 than AD-AMTS-3, whereas ADAMTS-3 mRNA was 5-fold higher than ADAMTS-2 mRNA in human cartilage. We propose that both ADAMTS-2 and ADAMTS-3 process procollagen II, but ADAMTS-3 is physiologically more relevant, given its preferred expression in cartilage. The findings provide an explanation for the sparing of cartilage in dermatosparaxis and, perhaps, for the relative sparing of some procollagen I-containing tissues.Collagens consist of the major structural proteins of the extracellular matrix (ECM) 1 and exist in both fibril-forming (e.g. collagens I-III, V, and XI) and nonfibrillar forms (1, 2). Molecules belonging to both categories are homotrimeric (e.g. collagen II) or heterotrimeric (e.g. collagen I) assemblies of specific ␣ chains, each the product of a single gene (1, 2). The molecular types of collagen, as well as the specific supramolecular aggregates they form, are often tissue-specific and provide a specialized function. For example, collagen I, the principal collagen of skin, is arranged in randomly oriented bundles in the dermis but in parallel bundles in tendons. Collagen II, a specific component of cartilage ECM, is arranged in an open meshwork that traps proteoglycans and facilitates resistance to compression. The synthesis, secretion, and assembly of collagens into specific supramolecular aggregates is a complex, multistep process (3, 4). Fibrillar collagens I-III are synthesized as a soluble procollagen monomer comprising a long triple helical "collagenous" region with smaller polypeptide extensions (propeptides) at the amino and carboxyl ends (4). Removal of the propeptides by specific enzymes, the N-and C-propeptidases (proteinases), is a prerequisite for the correct assembly of collagens I and II into growing fibrils (3, 4). The procollagen C-propeptidase is identical to bone morphogenetic protein-1 and processes all three of these fibrillar collagens (5). Biochemically distinct N-propeptidases with specificity for procollagens I and II or procollagen III are known (6). The bovine and human procollagen I N-propeptidases have been cloned (7,8). This enzyme (designated ADAMTS-2, EC 3.4.24.14), is a zinc...

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Assessment of procollagen processing defects by fibroblasts cultured in the presence of dextran sulphate

Cited by 51 publications

References 34 publications

In vitro effects of dextran sulfate sodium on a Caco-2 cell line and plausible mechanisms for dextran sulfate sodium-induced colitis

In vitro effects of dextran sulfate sodium on a Caco-2 cell line and plausible mechanisms for dextran sulfate sodium-induced colitis

Cloning and Characterization of ADAMTS-14, a Novel ADAMTS Displaying High Homology with ADAMTS-2 and ADAMTS-3

Procollagen II Amino Propeptide Processing by ADAMTS-3

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