High-performance liquid chromatographic determinations of disaccharides resulting from enzymatic degradation of isomeric chondroitin sulfates

Lee, George Jia-Long; Tieckelmann, Howard

doi:10.1016/0003-2697(79)90814-5

Cited by 82 publications

(7 citation statements)

References 20 publications

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“…The position of sulfation of the chondroitin sulfate chains was determined by high-performance liquid chromatography (34) of the unsaturated disaccharides produced by the action of chondroitinase ABC (35). The chromatography system utilized a LiChrosorb-NH2 column (36) and an acetate buffer system (37). Proteoglycan size was estimated by chromatography through Sepharose CL-2B as previously described (38), and the ability to aggregate was assessed by adding 2% hyaluronic acid (w/w) to the sample prior to chromatography.…”

Section: Methodsmentioning

confidence: 99%

The Structure and Abundance of Cartilage Proteoglycans during Early Development of the Human Fetus

Roughley

White

Glant³

1987

Pediatr Res

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ABSTRACT. Proteoglycan was isolated from the epiphyseal cartilage of the knee joint from human fetuses, ranging in age from 11 to 19 wk of gestation. The content of proteoglycan, per wet weight of cartilage, increased with gestational age, and structural changes were observed in the proteoglycan subunits, particularly with respect to hydrodynamic size and the position of sulfation of the chondroitin sulfate chains. While 6-sulfation remained fairly constant on about half the disaccharide residues during the age period examined, the proportion of nonsulfated residues decreased with gestational age and there was a corresponding increase in the proportion of 4-sulfated residues. Other structural parameters showed little change, and after 11 wk of gestation the majority of proteoglycan subunits were able to interact with hyaluronic acid to form aggregates. Link proteins were detected in the cartilage at all ages and their abundance increased with age relative to other cartilage proteins. They showed little change in structural heterogeneity, with the two larger molecular forms predominating. In contrast to the proteoglycans, cartilage proteins were in high abundance at 11 wk of gestation, but decreased considerably with time, although there was little change in the relative proportion of the majority of the components. (Pediatr Res 22: [409][410][411][412][413]1987) Abbreviation SDS, sodium dodecyl sulfateHyaline cartilage is best depicted as a solution of proteoglycan and matrix proteins entrapped within a network of collagen fibrils containing a sparse population of cells. The composition of the extracellular matrix is not constant throughout life, and this is particularly true for the proteoglycan, whose structure shows considerable variation. In the human the major changes after birth take place in the first two decades of life (I), representing the period of skeletal development. They are typified by a decrease in the size of the proteoglycan subunit, a decrease in both the size and number of chondroitin sulfate chains, and an increase in the size and number of keratan sulfate chains (2, 3). This latter change occurs concomitantly with a decrease in the number of 0-linked oligosaccharides (4, 5). Such changes in

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Section: Methodsmentioning

confidence: 99%

The Structure and Abundance of Cartilage Proteoglycans during Early Development of the Human Fetus

Roughley

White

Glant³

1987

Pediatr Res

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“…The ratio of chondroitin 4-and 6-sulfates was determined by disaccharide analysis. 30 The lipoprotein-proteoglycan complexes were also analyzed for free and esterified cholesterol, 31 phospholipids, 32 and total protein. 33 Triglycerides were determined by the Abbott V.P.…”

Section: Characterization Of Lipoprotein-proteoglycan Complexesmentioning

confidence: 99%

Lipoprotein-proteoglycan complexes from atherosclerotic lesions promote cholesteryl ester accumulation in human monocytes/macrophages.

Vijayagopal

Srinivasan

Radhakrishnamurthy

et al. 1992

Arterioscler Thromb

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Lipoprotein-proteoglycan complexes from human atherosclerotic lesions were studied to determine their ability to stimulate cholesteryl ester accumulation in human monocytes/macrophages. Complexes containing apolipoprotein (apo) B lipoproteins and proteoglycans were extracted from fatty streaks and fibrous plaque lesions of human aortas by extraction with 0.15 M NaCl. Fractionation of the complex with Bio-Gel A-50m yielded a single fraction from fatty streaks and two fractions from fibrous plaques. The complexes were further purified by antl-apo B affinity chromatography and analyzed for apolipoproteins, lipids, and glycosaminoglycans. Apo B was the only apolipoprotein present in the complexes. Although the complexes from fatty streaks and fibrous plaques contained varying proportions of hyaluronic acid, chondroitin 6-sulfate, and dermatan sulfate, heparin was present in only the fibrous plaque complexes. All three lipoprotein-proteogtycan complexes increased the rate of incorporation of [ M C]oleate into cholesteryl [ 14 C]oleate and stimulated cholesteryl ester accumulation in monocytes/macrophages. However, the complexes from fibrous plaques were more potent than those from fatty streaks in this regard. Cholesteryl ester synthesis that is mediated by the uptake of the complexes was dose dependent and showed apparent saturation, suggesting that cell surface binding may be required. Chloroquine, a lysosomotropic agent, inhibited cholesteryl ester synthesis that is induced by the complexes, indicating that lysosomal hydrolysis was essential. Cholesteryl ester synthesis that is mediated by the complexes was inhibited 70-79% by polyinosinic acid. Furthermore, excess unlabeled fibrous plaque complexes significantly inhibited the binding and interaalization of in vitro In vitro, low density lipoprotein (LDL) does not induce cholesteryl ester accumulation in macrophages from different sources, including human

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“…4P Heparitanase was prepared from Flavobacterium heparinum by the method of Silva et al 34 HS, in the amount of 300 jiig as uronic acid, was digested with 1 unit of heparitinase in 0.1 M acetate buffer, pH 7.0, for 20 h at 37°C. 35 By high performance liquid chromatography, 41 the degradation products of HS could be qualitatively measurable from the ultraviolet activity absorbance value by comparing them with similarly treated standard HS. The enzymatic activity was examined using standard C-4S, C-6S, HA and DS as substrates when the enzymes were of practical use.…”

Section: Preparation Of Enzymesmentioning

confidence: 99%

Distribution of acidic glycosaminoglycans, lipids and water in normal human cerebral arteries at various ages.

Murata¹

1985

Stroke

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SUMMARY Functional alterations in arterial acidic glycosaminoglycans (AGAG) may be related to the pathogenesis of some forms of cerebrovascular disease. We measured the AGAG, lipid and water content of human cerebral artery of 275 normal males at various ages. These measures were separately carried out in the main trunk and distal branches. The AGAG components were analyzed by an enzymatic assay method employing specific enzymes which digest AGAG to assess topographic change and aging variations. The total AGAG content was higher in the main cerebral artery than in the distal branches. The main AGAG component of the normal main cerebral artery was heparan sulfates (HS), constituting half the total AGAG, followed by moderate amounts of dermatan sulfate (DS), chondroitin-6-sulfate (C-6S) and chondroitin-4-sulfate (C-4S). Hyaluronic acid (HA) was a minor component and it was more prominent in young arteries. Heparin could be occasionally detected. With advancing age, the relative amounts of HS, HA, chondroitin and C-4S both in the main trunk and distal branches decreased but those of DS and C-6S increased. The total lipid, cholesterol ester and triglyceride content was greater in the main trunk than in the distal branches; the total lipid content increased with age. A possible function of the cerebral arterial AGAG is discussed with respect to change in lipid and water content according to topographic sites and aging.

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High-performance liquid chromatographic determinations of disaccharides resulting from enzymatic degradation of isomeric chondroitin sulfates

Cited by 82 publications

References 20 publications

The Structure and Abundance of Cartilage Proteoglycans during Early Development of the Human Fetus

The Structure and Abundance of Cartilage Proteoglycans during Early Development of the Human Fetus

Lipoprotein-proteoglycan complexes from atherosclerotic lesions promote cholesteryl ester accumulation in human monocytes/macrophages.

Distribution of acidic glycosaminoglycans, lipids and water in normal human cerebral arteries at various ages.

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