Immunochemical analysis of cartilage proteoglycans. Cross-reactivity of molecules isolated from different species

Wieslander, Jörgen

doi:10.1042/bj1990081

Cited by 15 publications

(4 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Comparison of the results indicates that the binding regions of cow (89%) and dog (78%) show a high degree of cross-reactivity with that of pig, with rat (37%) and human (22%) giving lower values and with rabbit (15%) being the lowest. Comparable results for antisera to proteoglycans from bovine nasal cartilage have been reported by Wieslander & Heinegard (1981). Discussion…”

Section: Fig 1 Titration Of (A) Anti-(link Protein) Antiserum and (supporting

confidence: 72%

Cartilage proteoglycan binding region and link protein. Radioimmunoassays and the detection of masked determinants in aggregates

Ratcliffe¹,

Hardingham²

1983

Biochemical Journal

View full text Add to dashboard Cite

Antibodies have been raised in rabbits to the hyaluronate-binding region and link-protein components of aggregated proteoglycans from pig laryngeal cartilage. The anti-(binding region) antibodies did not bind 125I-labelled link protein, nor was 125I-labelled binding region bound by the anti-(link protein) antibodies. The antisera were applied in sensitive inhibition radioimmunoassays to determine binding region and link protein in purified proteoglycan preparations. With intact proteoglycan aggregates, the antigenic sites of link protein, and to a lesser extent binding region, were masked. Heat treatment in the presence of sodium dodecyl sulphate (0.025%, w/v) was found to overcome this masking, thereby allowing the determination of link protein and binding region in aggregated proteoglycan preparations in pure and impure samples.

show abstract

Section: Fig 1 Titration Of (A) Anti-(link Protein) Antiserum and (supporting

confidence: 72%

Cartilage proteoglycan binding region and link protein. Radioimmunoassays and the detection of masked determinants in aggregates

Ratcliffe¹,

Hardingham²

1983

Biochemical Journal

View full text Add to dashboard Cite

show abstract

“…Furthermore, Triphaus et al (1980) found two metabolically distinct populations of proteoglycans in human xiphoid cartilage. Immunochemical studies indicated that cartilage extracts contain more than one population of proteoglycans (Wieslander & Heinegard, 1981), and in the present study we have purified two antigenically distinct populations of aggregating proteoglycans.…”

Section: Discussionmentioning

confidence: 53%

Separation and characterization of two populations of aggregating proteoglycans from cartilage

Heinegård¹,

Wieslander²,

Sheehan³

et al. 1985

Biochemical Journal

Self Cite

View full text Add to dashboard Cite

Intermediary gel immunoelectrophoresis was used to show that purified aggregating cartilage proteoglycans from 2-year-old steers contain two distinct populations of molecules and that only one of these is immunologically related to non-aggregating cartilage proteoglycans. The two types of aggregating proteoglycans were purified by density-gradient centrifugation in 3.5M-CsCl/4M-guanidinium chloride and separated by zonal rate centrifugation in sucrose gradients. The higher-buoyant-density faster-sedimenting proteoglycan represented 43% of the proteoglycans in the extract. It had a weight-average Mr of 3.5 X 10(6), did not contain a well-defined keratan sulphate-rich region, had a quantitatively dominant chondroitin sulphate-rich region and contained 5.9% protein and 23% hexosamine. The lower-buoyant-density, more slowly sedimenting, proteoglycan represented 15% of the proteoglycans in the extract. It had a weight-average Mr of 1.3 X 10(6), contained both the keratan sulphate-rich and the chondroitin sulphate-rich regions and contained 7.3% protein and 23% hexosamine. Each of the proteoglycan preparations showed only one band on agarose/polyacrylamide-gel electrophoresis. The larger proteoglycan had a lower mobility than the smaller. The distribution of chondroitin sulphate chains along the chondroitin sulphate-rich region was similar for the two types of proteoglycans. The somewhat larger chondroitin sulphate chains of the larger proteoglycan could not alone account for the larger size of the proteoglycan. Peptide patterns after trypsin digestion of the proteoglycans showed great similarities, although the presence of a few peptides not shared by both populations indicates that the core proteins are partially different.

show abstract

“…The large proteoglycan synthesized by arterial smooth muscle cells resembles the large proteoglycan population recently isolated from human aorta by Salisbury and Wagner (6) The smaller proteoglycan present in the medium contains dermatan sulfate. To some extent, this population resembles the small "ubiquitous" component produced by other mesenchymal ceils in vitro (49,50,52,53) and that recently isolated and characterized from cartilage (54). Several investigators have noticed the presence of dermatan sulfate proteoglycans in various tissue (55)(56)(57)(58)(59) including aorta (1,3,4,5,55).…”

Section: Discussionmentioning

confidence: 83%

Proteoglycans in primate arteries. III. Characterization of the proteoglycans synthesized by arterial smooth muscle cells in culture.

Wight¹,

Hascall²

1983

The Journal of Cell Biology

112

View full text Add to dashboard Cite

Near confluent monolayers of arterial smooth muscle cells derived from Macaca nemestrina were labeled with Na2[aSS]O4 and the newly synthesized proteoglycans present in the culture medium and cell layer were extracted with either 4 M guanidine HCI (dissociative solvent) or 0.5 M guanidine HCI (associative solvent) in the presence of protease inhibitors. The proteoglycans in both compartments were further purified by cesium chloride density gradient ultracentrifugation. Two size classes of proteoglycans were observed in the medium as determined by chromatography on Sepharose CL-2B. The large population ("Kay = 0.31) contained predominantly chondroitin sulfate chains with Mr = ~40,000. The smaller population (Kay = 0.61) contained dermatan sulfate chains of similar Mr (~40,000). When tested for their ability to aggregate, only proteoglycans in the large-sized population were able to aggregate. A chondroitin sulfate containing proteoglycan with identical properties was isolated from the cell layer. In addition, the cell layer contained a dermatan sulfate component which eluted later on Sepharose CL-2B (Kay = 0.78) than the dermatan sulfate proteoglycan present in the medium. Electron microscopy of the purified proteoglycans revealed a bottlebrush structure containing a central core averaging 140 nm in length with an average of 8 to 10 side projections. The length of the side projections varied but averaged between 70 and 75 nm. Similar bottlebrush structures were observed in the intercellular matrix of the smooth muscle cell cultures after staining with Safranin 0. This culture system provides a model to investigate parameters involved in the regulation of synthesis and degradation of arterial proteoglycans.Proteoglycans and their constituent glycosaminoglycans are present in blood vessel walls (1-14) and form important structural links between the fibrous components of the intercellular matrix (collagen and elastic fibers) and the arterial endothelial and smooth muscle cells (15). Recently, considerable attention has focused on these macromolecules since they have been shown to influence the structural integrity of the vascular wall and to affect several arterial properties such as viscoelasticity, permeability, lipid metabolism, hemostasis, and thrombosis (15,16). Furthermore, their participation in such cell processes as proliferation (17), migration (18), and adhesion (19) highlight their importance in many events that are common to both blood vessel development and disease.Although several studies have investigated changes in the glycosaminoglycan component of arterial proteoglycans during development and disease (10, 11,15,20), only recently have attempts been made to define the physical-chemical nature of entire proteoglycan molecules isolated from arteries. This information is important since it is thought that the structure of the whole molecule, and not merely the component parts, dictates the functional properties of the proteoglycans. Employing methods used to isolate proteoglycans from cartila...

show abstract

Immunochemical analysis of cartilage proteoglycans. Cross-reactivity of molecules isolated from different species

Cited by 15 publications

References 20 publications

Cartilage proteoglycan binding region and link protein. Radioimmunoassays and the detection of masked determinants in aggregates

Cartilage proteoglycan binding region and link protein. Radioimmunoassays and the detection of masked determinants in aggregates

Separation and characterization of two populations of aggregating proteoglycans from cartilage

Proteoglycans in primate arteries. III. Characterization of the proteoglycans synthesized by arterial smooth muscle cells in culture.

Contact Info

Product

Resources

About