Isolation of a unique collagenous fraction from limited pepsin digests of human placental tissue. Characterization of one of the constituent polypeptide chains.

Furuto, Donald K.; Miller, Edward J.

doi:10.1016/s0021-9258(19)86296-8

Cited by 108 publications

(23 citation statements)

References 24 publications

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“…Type VI collagen, best classed as a short-helix group 3 collagen molecule (Miller, 1985), is widely distributed among soft connective tissues, including aorta, placenta, uterus, skin, ligament, tendon and cornea (Chung et al, 1976;Furuto & Miller, 1980Jander et al, 1981Jander et al, , 1983Abedin et al, 1982;Odermatt et al, 1983;Furthmayr et al, 1983;Knight et al, 1984;Trueb & Bornstein, 1984;Gibson & Cleary, 1985;Engel et al, 1985;Ayad et al, 1985;Zimmermann et al, 1986). It is believed to be the major component of the microfibrillar structures seen in the matrices of these tissues (Von der Mark et al, 1984;Engel et al, 1985).…”

Section: Discussionmentioning

confidence: 99%

Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein

Eyre

Slayter

1987

Biochemical Journal

132

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The collagen framework of the intervertebral disc contains two major fibril-forming collagens, types I and II. Smaller amounts of other types of collagen are also present. On examination of the nature and distribution of these minor collagens within bovine disc tissue, type VI collagen was found to be unusually abundant. It accounted for about 20% of the total collagen in calf nucleus pulposus, and about 5% in the annulus fibrosus. It was discovered by serially digesting disc tissue with chondroitin ABC lyase and Streptomyces hyaluronidase that native covalent polymers of type VI collagen could be extracted. Electron micrographs of this material prepared by rotary shadowing revealed the characteristic dimensions of tetramers and double tetramers of type VI molecules, with their central rods and terminal globular domains. Molecular-sieve column chromatography on agarose under non-reducing non-denaturing conditions gave a series of protein peaks with molecular sizes equivalent to the tetramer, double tetramer and higher multimers. On SDS/polyacrylamide-gel electrophoresis after disulphide cleavage, these fractions of type VI collagen all showed a main band at Mr 140,000 and four lesser bands between Mr 180,000 and 240,000. On electrophoresis without disulphide cleavage in agarose/2.4% polyacrylamide only dimeric (six chains) and tetrameric (12 chains) forms of type VI molecules were present. The ability to extract all the type VI collagen of the tissue in 4 M-guanidinium chloride, and absence of aldehyde-mediated cross-linking residues on direct analysis, showed that, in contrast with most matrix collagens, type VI collagen does not function as a covalently cross-linked structural polymer.

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Section: Discussionmentioning

confidence: 99%

Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein

Eyre

Slayter

1987

Biochemical Journal

132

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“…These peptides also exhibit solubility properties similar to those of the G collagen described here but are distinguished from it by their lower molecular weights (33,000) and their occurrence as disulphide-bonded aggregates in the unreduced state (16) . Other short-chain collagens have also been isolated from pepsin digests of bovine (21) and human (13) placenta; these too are very soluble and disulphidelinked but have molecular weights (40,000) nearer those of the short-chain collagen synthesized by chrondrocytes in collagen gels.…”

Section: Collagen Types Deposited In the Cell Matrixmentioning

confidence: 99%

Effects of matrix macromolecules on chondrocyte gene expression: synthesis of a low molecular weight collagen species by cells cultured within collagen gels.

Gibson¹,

Schor²,

Grant³

1982

The Journal of Cell Biology

217

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Chick-embryo sternal chondrocytes have been cultured within three-dimensional Collagen gels as part of a study concerned with the effects of extracellular matrix macromolecules on chondrocyte gene expression . Data are presented indicating that chondrocytes cultured within such a collagenous environment synthesize significantly more of an hitherto unidentified, low molecular weight collagen species than do cells grown on plastic tissueculture dishes in the conventional manner . This low molecular weight collagen species contains noncollagenous domains (as indicated by its decreased molecular size after mild pepsin digestion), is distinct from the known collagen types (as judged by CNBr peptide analysis), and forms part of the insoluble collagenous matrix produced by the chondrocytes . Cells growing within the gel tend to form colonies consisting of a linear array of cells reminiscent of the cellular organization in growth cartilage.Although chondrocytes explanted to plastic tissue-culture dishes initially synthesize cartilage-specific macromolecules, it has proved difficult to maintain the expression of a differentiated phenotype during subsequent culture in vitro . The common observation is that chondrocytes in vitro tend to assume a fibroblastoid morphology with a concomitant switch from the predominant synthesis of type II to type I collagen (26).A number of culture parameters have been observed to affect chondrocyte behaviour in vitro . For example, Von der Mark et al. (39) have demonstrated that cell density and position within a colony influence the ability of chondrocytes to synthesize a cartilage-specific matrix. Cell-produced matrix macromolecules may play an important role in this regulatory process since the presence of exogenous collagen (23), proteoglycan (17, 18) and fibronectin (32, 40) have all been observed to influence the quantity of cartilage-specific components synthesized by chondrocytes in vitro . Indeed, the differentiation of a number of cell types including epidermal basal cells (29), hepatocytes (34), mammary epithelial cells (41) and fibroblasts (14) has been shown to be affected by culture on a collagen substratum.The present study was initiated to investigate further the effects of a collagen substratum on the synthesis of different

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“…YPE VI collagen is widely distributed throughout connective tissues (l 3, 24). It has been isolated from aorta and placenta in the form of high molecular weight, pepsin-resistant fragments composed of 50-80-kD polypeptides (3,8,9,19,13,14,16,19,21). It has also been isolated from these tissues by guanidine hydrochloride extraction as a protein composed of polypeptides ranging from 140 kD (10,15) up to 190 kD (21).…”

mentioning

confidence: 99%

Molecular assembly, secretion, and matrix deposition of type VI collagen.

Engvall¹,

Hessle²,

Klier³

1986

The Journal of Cell Biology

245

160

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Abstract. Monoclonal antibodies reactive with the tissue form of type VI collagen were used to isolate the type VI collagen polypeptides from cultured fibroblasts and muscle cells. Two [35S]methionine-labeled polypeptides of 260 and 140 kD were found intracellularly, in the medium, and in the extracellular matrix of metabolically labeled cells. These polypeptides were disulfide cross-linked into very large complexes. The 260-and 140-kD polypeptides were intimately associated and could not be separated from each other by reduction without denaturation. In the absence of ascorbic acid, both polypeptides accumulated inside the cell, and their amounts in the medium and in the matrix were decreased. These results suggest that both the 260-and the 140-kD polypeptides are integral parts of the type VI collagen molecule. Examination of type VI collagen isolated from the intracellular pool by electron microscopy after rotary shadowing revealed structures corresponding to different stages of assembly of type VI collagen. Based on these images, a sequence for the intracellular assembly of type VI collagen could be discerned. Type VI collagen monomers are ~ 125 nm long and are composed of two globules separated by a thin strand. The monomers assemble into dimers and tetramers by lateral association. Only tetramers were present in culture media, whereas both tetramers and multimers were found in extracellular matrix extracts. The multimers appeared to have assembled from tetramers by end-to-end association into filaments that had prominent knobs and a periodicity of-110 nm. These results show that, unlike other collagens, type VI collagen is assembled into tetramers before it is secreted from the cells, and they also suggest an extracellular aggregation mechanism that appears to be unique to this collagen.T YPE VI collagen is widely distributed throughout connective tissues (l 3, 24). It has been isolated from aorta and placenta in the form of high molecular weight, pepsin-resistant fragments composed of 50-80-kD polypeptides (3,8,9,19,13,14,16,19,21). It has also been isolated from these tissues by guanidine hydrochloride extraction as a protein composed of polypeptides ranging from 140 kD (10, 15) up to 190 kD (21). One outstanding feature of both the pepsin fragments and the larger forms of type VI collagen is the extensive interchain disulfide cross-linking. It appears from chemical and morphological analyses of type VI collagen that it contains relatively short triple-helical domains, contributing one-third or less to the total mass of the protein (7,10,13,15,19).The molecular architecture (2, 7, 13, 15), the large amount of globular structure, and the extensive disulfide cross-linking in type VI collagen suggest that it is unique among collagens, but little is known about its biosynthesis, processing, and biological function. Type VI collagen is synthesized by fibroblasts and muscle cells in culture and is deposited into their extracellular matrix (l l, 13, 22, 24). The type VI collagen immunoreactivity in cell cultur...

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Isolation of a unique collagenous fraction from limited pepsin digests of human placental tissue. Characterization of one of the constituent polypeptide chains.

Cited by 108 publications

References 24 publications

Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein

Type VI collagen of the intervertebral disc. Biochemical and electron-microscopic characterization of the native protein

Effects of matrix macromolecules on chondrocyte gene expression: synthesis of a low molecular weight collagen species by cells cultured within collagen gels.

Molecular assembly, secretion, and matrix deposition of type VI collagen.

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