070 Isolation and characterization of heparan sulphate proteoglycan from human articular cartilage

1992 ◽  
Vol 343 (1) ◽  
pp. 128-128
Author(s):  
G. St�cker ◽  
D. -C. Fischer ◽  
S. Handt ◽  
H. Greiling ◽  
H. -D. Haubeck
Keyword(s):  
Articular Cartilage ◽  
Heparan Sulphate ◽  
Human Articular Cartilage ◽  
Heparan Sulphate Proteoglycan ◽  
Sulphate Proteoglycan ◽  
Isolation And Characterization

scholarly journals Proteoglycan synthesis in human and murine haematopoietic progenitor cell lines: isolation and characterization of a heparan sulphate proteoglycan as a major proteoglycan from the human haematopoietic cell line TF-1

1996 ◽  
Vol 317 (1) ◽  
pp. 203-212 ◽  
Author(s):  
Georg STÖCKER ◽  
Zofia DRZENIEK ◽  
Ursula JUST ◽  
Wolfram OSTERTAG ◽  
Barbara SIEBERTZ ◽  
...  
Keyword(s):  
Cell Line ◽  
Stromal Cells ◽  
Progenitor Cell ◽  
Heparan Sulphate ◽  
Proteoglycan Synthesis ◽  
Heparan Sulphate Proteoglycan ◽  
Haematopoietic Progenitor Cell ◽  
Sulphate Proteoglycan ◽  
Isolation And Characterization

Proteoglycans of bone-marrow stromal cells and their extracellular matrix are important components of the microenvironment of haematopoietic tissues. Proteoglycans might also be involved in the interaction of haematopoietic stem and stromal cells. Recently, several studies have been reported on the proteoglycan synthesis of stromal cells, but little is known about the proteoglycan synthesis of haematopoietic stem or progenitor cells. Here we report on the isolation and characterization of proteoglycans from two haematopoietic progenitor cell lines, the murine FDCP-Mix A4 and the human TF-1 cell line. Proteoglycans were isolated from metabolically labelled cells and purified by several chromatographic steps, including anion-exchange and size-exclusion chromatography. Biochemical characterization was performed by electrophoresis or gel-filtration chromatography before and after digestion with glycosaminoglycan-specific enzymes or HNO2 treatment. Whereas FDCP-Mix A4 cells synthesize a homogeneous chondroitin 4-sulphate proteoglycan, isolation and characterization of proteoglycans from the human cell line TF-1 revealed, that TF-1 cells synthesize, in addition to a chondroitin sulphate proteoglycan, a heparan sulphate proteoglycan as major proteoglycan. For this heparan sulphate proteoglycan a core protein size of approx. 59 kDa was determined. Immunochemical analysis of this heparan sulphate proteoglycan revealed that it is not related to the syndecan family nor to glypican.

scholarly journals Purification and characterization of heparan sulphate proteoglycan from bovine brain

1999 ◽  
Vol 344 (3) ◽  
pp. 723 ◽  
Author(s):  
Youmie PARK ◽  
Guyong YU ◽  
Nur Sibel GUNAY ◽  
Robert J. LINHARDT
Keyword(s):  
Heparan Sulphate ◽  
Bovine Brain ◽  
Heparan Sulphate Proteoglycan ◽  
Purification And Characterization ◽  
Sulphate Proteoglycan

Characterization of an antiserum to a basement membrane heparan sulphate proteoglycan

1987 ◽  
Vol 15 (6) ◽  
pp. 1076-1077 ◽  
Author(s):  
MICHAEL A. F. DAVIS ◽  
WALTER T. GIBSON ◽  
HELEN J. MINTER ◽  
COLIN G. SMITH ◽  
JOHN R. COUCHMAN
Keyword(s):  
Basement Membrane ◽  
Heparan Sulphate ◽  
Heparan Sulphate Proteoglycan ◽  
Sulphate Proteoglycan

scholarly journals Structural differences and the presence of unsubstituted amino groups in heparan sulphates from different tissues and species

1997 ◽  
Vol 322 (2) ◽  
pp. 499-506 ◽  
Author(s):  
Toshihiko TOIDA ◽  
Hisao YOSHIDA ◽  
Hidenao TOYODA ◽  
Ichiro KOSHIISHI ◽  
Toshio IMANARI ◽  
...  
Keyword(s):  
Acid Content ◽  
Heparan Sulphate ◽  
Kidney Medulla ◽  
Isolation And Characterization ◽  
Iduronic Acid ◽  
Structural Differences ◽  
Liver And Kidney ◽  
High Degree ◽  
Different Tissues

This study presents a comparison of heparan sulphate chains isolated from various porcine and bovine tissues. 1H-NMR spectroscopy (500 MHz) was applied for structural and compositional studies on intact heparan sulphate chains. After enzymic digestion of heparan sulphate using heparin lyase I (EC 4.2.2.7) II and III (EC 4.2.2.8), the compositions of unsaturated disaccharides obtained were determined by analytical capillary electrophoresis. Correlations between the N-sulphated glucosamine residues and O-sulphation and between iduronic acid content and total sulphation were discovered using the data obtained by NMR and disaccharide analysis. Heparan sulphate chains could be classified into two groups based on the sulphation degree and the iduronic acid content. Heparan sulphate chains with a high degree of sulphation possessed also a significant number of iduronic acid residues and were isolated exclusively from porcine brain, liver and kidney medulla. The presence and amount of N-unsubstituted glucosamine residues (GlcNp) was established in all of the heparan sulphates examined. The structural context in which this residue occurs was demonstrated to be: high sulphation domain → 4)-β-d-GlcAp-(1 → 4)-α-d-GlcNp-(1 → 4)-β-d-GlcAp-(1 → low sulphation domain (where GlcNp is 2-amino-2-deoxyglucopyranose, and GlcAp is glucopyranosyluronic acid), based on the isolation and characterization of a novel, heparin lyase III-derived, GlcNp containing tetrasaccharide and hexasaccharide. The results presented suggest that structural differences may play a role in important biological events controlled by heparan sulphate in different tissues.

Thyroid hormone excess stimulates the synthesis of proteoglycan in human skin fibroblasts in culture

1990 ◽  
Vol 123 (5) ◽  
pp. 541-549 ◽  
Author(s):  
Yoshimasa Shishiba ◽  
Yasuhiro Takeuchi ◽  
Noriko Yokoi ◽  
Yasunori Ozawa ◽  
Taeko Shimizu
Keyword(s):  
Thyroid Hormone ◽  
Human Skin ◽  
Specific Activity ◽  
Heparan Sulphate ◽  
Skin Fibroblasts ◽  
Proteoglycan Synthesis ◽  
Heparan Sulphate Proteoglycan ◽  
Human Skin Fibroblasts ◽  
Dermatan Sulphate ◽  
Sulphate Proteoglycan

Abstract We previously demonstrated that proteoglycan accumulated in the affected skin of circumscribed pretibial myxedema of Graves' disease. As an underlying mechanism responsible for the accumulation, we sought to determine whether excess thyroid hormone was partially responsible for the increase in proteoglycan synthesis. Human skin fibroblasts were cultured in Ham's F-10 medium containing 1% Nutridoma with graded doses of T3 (0.184 × 10−9 to 46 × 10−9 mol/l) and were labelled with [35S]sulphate and [3H]glucosamine. Proteoglycans were purified by Sephadex G-50, Q-Sepharose chromatography with NaCl-gradient and Sepharose CL-6B chromatography. 35S and 3H incorporated into dermatan sulphate proteoglycan and heparan sulphate proteoglycan and 3H incorporated into hyaluronan were measured. 35S and 3H incorporation into dermatan sulphate proteoglycan was minimum at a T3 concentration of 0.184 × 10−9 mol/l, and increased with increasing doses of T3 up to 46 × 10−9 mol/l. 35S and 3H incorporation into heparan sulphate proteoglycan also increased with increasingdoses of T3. 3H incorporation into hyaluronan was not influenced at all by T3. The increased incorporation of 35S into proteoglycan in high-T3 culture reflects the increased synthesis of proteoglycan because 1. the extent of sulphation of disaccharides examined by thin-layer chromatography was not altered by T3; 2. the specific activity of [35S]sulphate was not influenced by T3, and 3. T3 did not decrease the degradation rate of cell-associated proteoglycan.

Sign in / Sign up

Export Citation Format

Share Document