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Structure and interactions of heparan sulfate proteoglycans from a mouse tumor basement membrane.
- Source :
-
European journal of biochemistry [Eur J Biochem] 1984 Aug 15; Vol. 143 (1), pp. 145-57. - Publication Year :
- 1984
-
Abstract
- Various forms of heparan sulfate proteoglycan were solubilized from the mouse Engelbreth-Holm-Swarm (EHS) sarcoma by extraction with 0.5 M NaCl, collagenase digestion and extraction with 4 M guanidine. They could be separated into high (greater than or equal to 1.65 g/ml) and low (1.38 g/ml) buoyant density variants. The high-density form from the NaCl extract and collagenase digest had Mr = 130000 and So20,W = 4.5 S and contained 4-10% protein, indicating Mr = 5 000-12 000 for the protein core. This proteoglycan exhibited polydispersity as shown by rotary shadowing electron microscopy and ultracentrifugation. An average molecule consisted of four heparan sulfate chains (Mr = 29 000) each with a length of 32 +/- 10 nm. The low-density form (Mr about 400 000) could not be completely purified and contained about 50% protein. As shown by radioimmunoassay, the various proteoglycans shared similar protein cores. Labeling of the tumor in vivo or in vitro demonstrated preferential incorporation of radioactive sulfate in the high-density form. The high-density proteoglycan interacted in affinity chromatography by virtue of its heparan sulfate chains with laminin, fibronectin, the globular domain NC1 and the triple helix of collagen IV. These interactions were abolished at moderate concentrations of NaCl (0.1-0.2 M) and in the presence of heparin, chondroitin sulfate or dextran sulfate. Interactions with the globule NC1 could also be demonstrated by velocity band centrifugation in sucrose gradients and a binding constant of about 10(6) M-1 was derived.
- Subjects :
- Animals
Basement Membrane analysis
Centrifugation, Density Gradient
Chromatography, Affinity
Chromatography, DEAE-Cellulose
Chromatography, Gel
Heparan Sulfate Proteoglycans
Mice
Mice, Inbred C57BL
Microscopy, Electron
Osmolar Concentration
Ultracentrifugation
Chondroitin Sulfate Proteoglycans analysis
Glycosaminoglycans analysis
Heparitin Sulfate analysis
Proteoglycans analysis
Sarcoma, Experimental analysis
Subjects
Details
- Language :
- English
- ISSN :
- 0014-2956
- Volume :
- 143
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- European journal of biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 6236080
- Full Text :
- https://doi.org/10.1111/j.1432-1033.1984.tb08353.x