[2] Isolation and characterization of collagens and procollagens

Furuto, Donald K.; Miller, Edward J.

doi:10.1016/0076-6879(87)44171-2

Cited by 20 publications

(5 citation statements)

References 57 publications

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“…These collagen types are present in the blood vessel wall or can be isolated from various tissues such as the human placenta, skin, or various tendons. 45,46 Several types of collagen were compared in an intensive study by Favaloro 41 for their ability to discriminate between different vWD subtypes when used in a collagen-binding ELISA. In this study, the collagen type and source were found to be crucial, but the effectiveness of the assay depended on the concentration used for immobilization.…”

Section: Human Vwf: Collagen-binding Activitymentioning

confidence: 99%

Comparative Study on Collagen-Binding Enzyme-Linked Immunosorbent Assay and Ristocetin Cofactor Activity Assays for Detection of Functional Activity of von Willebrand Factor

Turecek¹,

Siekmann²,

Schwarz³

2002

Semin Thromb Hemost

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For more than two decades, the ristocetin cofactor (RCo) assay, which measures the von Willebrand factor (vWF)-mediated agglutination of platelets in the presence of the antibiotic ristocetin, has been the most common method for measuring the functional activity of vWF. There is, however, general agreement among clinical analysts that this method has major practical disadvantages in performance and reproducibility. Today, collagen-binding assays (CBA) based on the enzyme-linked immunosorbent assay (ELISA) technique that measure the interaction of vWF and collagen are an alternative analytic procedure based on a more physiological function than that of the RCo procedure. We used both assay systems in a comparative study to assess the functional activity of vWF in plasma as well as in therapeutic preparations. We measured RCo activities of plasma from healthy donors and patients with different types of von Willebrand disease (vWD) and of vWF as a drug substance in factor (F) VIII/vWF concentrates using both the aggregometric and the macroscopic methods. In addition, we measured collagen-binding activity (vWF:CB) using a recently developed commercially available CBA system. To investigate the relation between the structure and the functional activity of vWF, we isolated vWF species with different numbers of multimers from FVIII/vWF concentrates by affinity chromatography on immobilized heparin. The vWF:RCo and vWF:CB of the different fractions were measured, and the multimeric structure of vWF was analyzed by sodium dodecyl sulfate (SDS) agarose gel electrophoresis. (vWF:CB and vWF:RCo are part of the nomenclature proposed by the International Society on Thrombosis and Hemostasis Scientific and Standardization Committee [ISTH SSC] subcommittee on von Willebrand factor, in Maastricht, Germany, June 16, 2000.) Measurement of functional vWF activity by CBA can be carried out with substantially higher interassay reproducibility than can measurement of RCo. Both assay systems can be used for diagnosis and subtyping of vWD, but CBA is more sensitive than either of the two RCo methods. The analysis of vWF multimers in the different fractions obtained by affinity chromatography on heparin Sepharose showed that the activity measured both with RCo assay and CBA correlated with the degree of multimerization. Our results suggest that measurement of the functional activity of vWF by the RCo procedure can be replaced by the more reliable CBA, reflecting the physiological hemostatic activity of vWF. The CBA method appears not only to be more sensitive and easier to carry out than the RCo method is but also to have a higher reproducibility and allow better standardization.

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Section: Human Vwf: Collagen-binding Activitymentioning

confidence: 99%

Comparative Study on Collagen-Binding Enzyme-Linked Immunosorbent Assay and Ristocetin Cofactor Activity Assays for Detection of Functional Activity of von Willebrand Factor

Turecek¹,

Siekmann²,

Schwarz³

2002

Semin Thromb Hemost

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“…The possibility that autoimmunity to types II and XI collagen might contribute to the pathogenesis of human rheumatic disease has been suggested by studies demonstrating antibodies to these collagens in the sera of patients with rheumatoid arthritis and RP [20,21]. Of the three cartilage‐specific collagens isolated, type II is the most abundant, constituting 80–90% of the total collagen content of the cartilage compared to types XI and IX collagen, which each constitute 5–10% of cartilage collagen [22,23].…”

Section: Discussionmentioning

confidence: 99%

Vaccination with collagen-pulsed dendritic cells prevents the onset and reduces the disease severity in the mouse model of spontaneous polychondritis

Sidhu

Griffiths

Bradley

2009

Clinical and Experimental Immunology

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SummaryImmature dendritic cells (iDCs) have a tolerogenic potential due to low expression of important co-stimulatory cell surface molecules required for antigen presentation and induction of an effective immune response. We report here that injection of iDCs pulsed with chick type II collagen (CII) delayed the onset significantly and suppressed the severity of spontaneous polychondritis (SP) in the human leucocyte antigen (HLA)-DQ6ab8ab transgenic mouse model. Bone marrow-derived iDCs were pulsed in vitro with CII and transferred into 6-week-old HLA-DQ6ab8ab transgenic mice. Mice receiving CII-pulsed iDCs did not display any clinical signs of disease until 5·5 months of age, indicating the ability of the DC vaccine to delay significantly the onset of SP. Control groups receiving unpulsed iDCs or phosphate-buffered saline (PBS) developed polyarthritis at 3·5 months, as we have reported previously. The severity and incidence of disease was reduced in mice injected with CII-pulsed iDCs. Proinflammatory cytokines were in low to undetectable levels in the serum and tissue in the CII-pulsed iDC mice, correlating with the protection. This is the first evidence of iDC therapy controlling SP and suggests that iDC vaccination may provide a tool to reducing clinical manifestations in human inflammatory autoimmune disease such as relapsing polychondritis and rheumatoid arthritis.

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“…Procollagen is defined as the soluble biosynthetic precursor of collagen. Cell culture mediumoften contains intact procollagen and intermediate forms of the molecule generated in the processing to the final product, collagen (12,33,42). In this report, we purified GNCP, a type of procollagen, from a culture mediumof mouse cells using heparin affinity chromatography.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Cell-Adhesive Proteins Secreted by Cell Lines Growing in Protein- and Lipid-Free Synthetic Medium. Mouse LP3 Cells Secrete a Procollagen Molecule with Potent Cell-Attachment Activity.

Morita

Yasumitsu

Watanabe³

et al. 1993

Cell Struct. Funct.

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ABSTRACT. Wehave previously shown that mouse L-P3 cells secrete fibronectin and a novel protein, gelatin-non-binding and heparin-binding cell-adhesive protein (GNCP). Here, we screened and characterized cell-adhesive proteins in the conditioned media (CM's) of a series of cell lines growing in a protein-and lipidfree synthetic medium (P3 cell lines). Although cell-attachment activity of the CM's ranged from undetectable to highly significant, fractionation with affinity columns revealed the presence of significant cell-attachment activity in all CM's. Using cell-attachment assay and immunoassay on blotted filters, various cell-adhesive proteins were detected in the CM's, and most of them were identified as fibronectin, laminin, vitronectin, and collagen. GNCP-like proteins were detected in the CM's ofHeLa P3, JTC-16 P3, L P3, and JTC-12 P3. There was no relationship between the origin of the cell lines and the cell-adhesive proteins secreted. GNCPpurified from L P3-CM was separated into 120-, 140-, 150-, and 160-kDa proteins on SDS-PAGE,which were judged to be a type of mouse type I procollagen from the following results: (1) they were digested by collagenase, (2) pepsin treatment converted the 150-and 160-kDa proteins into 120-and 140-kDa proteins, (3) they were recognized by anti-type I collagen antiserum, and (4) amino-terminal sequence of the pepsin-digested 140-kDa protein had significant homology with type I collagen. GNCPshowed half-maximum activity of cell attachment at 0.03^g/ml, indicating that GNCPwas a cell-adhesive protein with an extremely high specific activity compared to other knowncell-adhesive proteins.Cell-adhesive proteins are a general term of proteins capable of mediating adhesion of cells to the substrate. The most representative cell-adhesive proteins are fibronectin (18, 35,48,49), laminin (31), and collagen (13, 25), which are well-known as the major components of extracellular matrix. Fibronectin is also a component of blood plasma. Vitronectin is a potent cell-adhesive protein present in blood plasma (2), and is also found on cell surfaces and in tissues (16). These cell-adhesive proteins are involved in various biological processes, such as cell proliferation, migration, differentiation, development, and tumorigenesis and metastasis (29, 30, 39). can grow in a completely protein-and lipid-free synthetic medium. Somereports described the secretion of growth factors by these cell lines; that is, DNAsynthesis-stimulating activity and/or transforming growth factor (TGF)-like activity (36), and colony-stimulating fac- tor (46, 47). Other than these growth factors, P3 cell lines, which attach, spread, and grow on the surface of culture dishes, can be expected to produce some molecules that promote cell adhesion as a fundamental component for their own growth. We have reported that mouse L P3 cells, one of the P3 cell lines, secrete two types of cell-adhesive proteins (34). They were separated from each other by affinity chromatography on gelatin-and heparin-Sepharose columns. One was a ge...

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[2] Isolation and characterization of collagens and procollagens

Cited by 20 publications

References 57 publications

Comparative Study on Collagen-Binding Enzyme-Linked Immunosorbent Assay and Ristocetin Cofactor Activity Assays for Detection of Functional Activity of von Willebrand Factor

Comparative Study on Collagen-Binding Enzyme-Linked Immunosorbent Assay and Ristocetin Cofactor Activity Assays for Detection of Functional Activity of von Willebrand Factor

Vaccination with collagen-pulsed dendritic cells prevents the onset and reduces the disease severity in the mouse model of spontaneous polychondritis

Characterization of Cell-Adhesive Proteins Secreted by Cell Lines Growing in Protein- and Lipid-Free Synthetic Medium. Mouse LP3 Cells Secrete a Procollagen Molecule with Potent Cell-Attachment Activity.

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