Extraction and purification of proteoglycans from various types of connective tissue

Antonopoulos, C.A.; Axelsson, Inge; Heinegård, Dick; Gardell, S.

doi:10.1016/0304-4165(74)90341-9

Cited by 130 publications

(60 citation statements)

References 16 publications

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“…Such degradation appears highly improbable, in view of the following facts. (a) The isolation procedure employed in the present investigation has been extensively, and successfully, applied to the isolation of intact proteoglycans from several types of tissues (Antonopoulos et al, 1974); although this procedure differs considerably from that used in our previous study (Lindahl, 1970), similar low xylose, galactose and serine contents were recorded for the isolated heparin products, irrespective of the preparation procedure used. migrating like the charged disaccharide, glucuronosyl-anhydromannitol.…”

Section: Macromolecular Properties Of Heparin and Dermatan Sulphate Fmentioning

confidence: 94%

Macromolecular properties and end-group analysis of heparin isolated from bovine liver capsule

Jansson¹,

Ogren²,

Lindahl³

1975

Biochemical Journal

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Glycosaminoglycans were extracted from bovine liver capsule with 4M-guanidinium chloride, resulting in solubilization of approx. 90% of the total uronic acid-containing polysaccharide of the tissue. The extracted polysaccharide was purified and fractionated by anion-exchange chromatography on DEAE-cellulose, density-gradient ultracentrifugation in CsCl and finally gel chromatography on Sepharose 4B. By using these procedures, the two major polysaccharide components, dermatan sulphate and heparin, which constituted 55 and 30 % respectively of the total glycosaminoglycan content of the tissue, were separated from each other. Analysis of the macromolecular properties of the two polysaccharides showed that heparin existed exclusively as single polysaccharide chains, whereas dermatan sulphate occurred largely as a proteoglycan (protein content, 74% dry wt.). The purified heparin preparation was subjected to sedimentationequilibrium ultracentrifugation, indicating a molecular weight of 8800. Analysis for neutral sugars (by g.l.c.) showed 0.1 residue of xylose and 0.2 residue of galactose/ polysaccharide chain; serine amounted to 0.3 residue/polysaccharide chain. Reduction of the heparin with NaB3H4 resulted in incorporation of 3H, approximately corresponding to one reducible group/polysaccharide chain. The 3H-labelled sugar residue was liberated by a combination of acid hydrolysis and deaminative cleavage of the polysaccharide with HNO2; it was subsequently identified as an aldonic acid by paper electrophoresis. Most of the heparin chains thus contained a uronic acid residue in reducing position. It is suggested that heparin isolated from bovine liver capsule is a degradation product released from larger molecules by an endo-glycuronidase.

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Section: Macromolecular Properties Of Heparin and Dermatan Sulphate Fmentioning

confidence: 94%

Macromolecular properties and end-group analysis of heparin isolated from bovine liver capsule

Jansson¹,

Ogren²,

Lindahl³

1975

Biochemical Journal

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“…The extracellular matrix of the normal corneal stroma consists primarily of type I collagen and two proteoglycans: a chondroitin-dermatan sulfate proteoglycan and a keratan sulfate proteoglycan (4)(5)(6)(7)(8)(9). Because most of the sulfated carbohydrate in the cornea is contained in these two proteoglycans, macular corneal dystrophy may involve alterations in one or both of these proteoglycans.…”

mentioning

confidence: 99%

Macular corneal dystrophy: failure to synthesize a mature keratan sulfate proteoglycan.

Hassell¹,

Newsome²,

Krachmer³

et al. 1980

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

151

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Corneal specimens obtained during surgery from patients with macular corneal dystrophy and obtained at autopsy from control eyes were incubated in a medium containing radioactive precursors of glycoproteins and proteoglycans. Biosyntheticallyradiolabeled material was extracted and characterized by using molecular sieve chromatography and specific enzymes. Cells in control corneas synthesized both a chondroitin sulfate proteoglycan and a keratan sulfate proteoglycan similar to those present in monkey and bovine corneas. Cells in macular corneas synthesized a normal chondroitin sulfate proteoglycan but did not synthesize either keratan sulfate or a mature keratan sulfate proteoglycan. Instead, macular corneas synthesized a glycoprotein with unusually large oligosaccharide side chains. This glycoprotein was not detected in normal corneas and is slightly smaller than normal keratan sulfate prot can. The failure to synthesize a mature keratan sulfate proteoglycan may produce corneal opacity and result in blindness. Because of evidence indicating that the corneal keratan sulfate proteoglycan is normally synthesized through a glycoprotein intermediate [Hart, G. W. & Lennarz, W. (1978) J. Biol. Chem. 253,5795-5801, macular corneal dystrophy may be a defect in glycoprotein processing. Macular, lattice, and granular corneal dystrophies are characterized by specific abnormalities of the corneal stroma and result in loss of vision (1). Macular corneal dystrophy is characterized clinically by recessive inheritance, a general diffuse haze throughout the corneal stroma, and the accumulation of irregularly shaped deposits which tend to be anterior in the central region of the stroma. Histologically, the deposits are primarily extracellular and stain positively with Alcian blue and the periodic acid-Schiff reagent (2, 3). These observations suggest that the deposits may be glycosaminoglycans and oligosaccharides accumulating in the stromal extracellular matrix.The extracellular matrix of the normal corneal stroma consists primarily of type I collagen and two proteoglycans: a chondroitin-dermatan sulfate proteoglycan and a keratan sulfate proteoglycan (4-9). Because most of the sulfated carbohydrate in the cornea is contained in these two proteoglycans, macular corneal dystrophy may involve alterations in one or both of these proteoglycans. Recently, an organ culture system utilizing monkey corneas has been developed that allows the biosynthesis of radiolabeled corneal stromal proteoglycans in vItro (9). This system involves incubating intact corneas in a tissue culture medium containing radioactive precursors. Analyses of the radiolabeled proteoglycans synthesized during organ culture indicated that they were representative of the proteoglycans that accumulate in the stromal extracellular matrix.The corneal cloudiness in individuals with macular corneal MATERIALS AND METHODS Organ Culture Procedures. Corneas excised with a 7-to 8-mm trephine blade were obtained from human eyes at keratoplasty or from control donor eyes a...

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“…bovine gastrointestinal tract; proteodermatan sulfate; endo-/3-xylosidase; extracellular matrix; glycosaminoglycan Proteoglycans (PGs) are important structural macromolecules in the extracellular matrix (Antonopoulos et al 1974;Vogel and Fisher 1986). Interest in PGs…”

Section: Kawasaki H Comparative Studies On Proteodermatan Sulfate Omentioning

confidence: 99%

“…Dermatan sulfate chains differed from each other to a slight degree with respect to chain length and sulfation, that from the esophagus being the largest. These differences in proteodermatan sulfate structure seem to reflect the organ specificities of the gastrointestinal tract.bovine gastrointestinal tract; proteodermatan sulfate; endo-/3-xylosidase; extracellular matrix; glycosaminoglycan Proteoglycans (PGs) are important structural macromolecules in the extracellular matrix (Antonopoulos et al 1974;Vogel and Fisher 1986). Interest in PGs…”

mentioning

confidence: 99%

Comparative Studies on Proteodermatan Sulfate of Bovine Gastrointestinal Tract.

Kawasaki

1993

Tohoku J. Exp. Med.

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KAWASAKI, H. Comparative Studies on Proteodermatan Sulfate of BovineGastrointestinal Tract. Tohoku J. Exp. Med., 1993, 171(3), 255-266 Proteodermatan sulfate was extracted from four areas of the bovine gastrointestinal tract (esophagus, stomach, small intestine and colon) with 4 M guanidine-HC1 and then purified by ion-exchange and gel filtration chromatography. Dermatan sulfate chains which made up proteodermatan sulfate from each area were separately prepared by Pronase P and endo-f3-xylosidase digestion. The properties of the proteodermatan sulfate and dermatan sulfate chains were compared using electrophoresis and high-performance liquid chromatography. The molecular size of proteodermatan sulfate purified from each area was estimated to be greater than 8 X 105, and the molecular sizes of dermatan sulfate chains from esophagus, stomach, small intestine, and colon were 27,000, 24,500, 21,000, and 21,500, respectively. The dermatan sulfate chains from the esophagus were slightly undersulfated in comparison with the others. These results show that the molecular sizes of proteodermatan sulfate from different regions of the gastrointestinal tract are similar to each other, but are larger than those from other tissues. Dermatan sulfate chains differed from each other to a slight degree with respect to chain length and sulfation, that from the esophagus being the largest. These differences in proteodermatan sulfate structure seem to reflect the organ specificities of the gastrointestinal tract.bovine gastrointestinal tract; proteodermatan sulfate; endo-/3-xylosidase; extracellular matrix; glycosaminoglycan Proteoglycans (PGs) are important structural macromolecules in the extracellular matrix (Antonopoulos et al. 1974;Vogel and Fisher 1986). Interest in PGs

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Extraction and purification of proteoglycans from various types of connective tissue

Cited by 130 publications

References 16 publications

Macromolecular properties and end-group analysis of heparin isolated from bovine liver capsule

Macromolecular properties and end-group analysis of heparin isolated from bovine liver capsule

Macular corneal dystrophy: failure to synthesize a mature keratan sulfate proteoglycan.

Comparative Studies on Proteodermatan Sulfate of Bovine Gastrointestinal Tract.

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