Versican V2 is a major extracellular matrix component of the mature bovine brain.
We have isolated and characterized the proteoglycan isoforms of versican from bovine brain extracts. Our approach included (i) cDNA cloning and sequencing of the entire open reading frame encoding the bovine versican splice variants; (ii) preparation of antibodies against bovine versican using recombinant core protein fragments and synthetic peptides; (iii) isolation of versican isoforms by ammonium sulfate precipitation followed by anion exchange and hyaluronan affinity chromatography; and (iv) characterization by SDS-polyacrylamide gel electrophoresis and Coomassie Blue staining or immunoblotting. Our results demonstrate that versican V2 is, together with brevican, a major component of the mature brain extracellular matrix. Versicans V0 and V1 are only present in relatively small amounts. Versican V2 migrates after chondroitinase ABC digestion with an apparent molecular mass of about 400 kDa, whereas it barely enters a 4-15% polyacrylamide gel without the enzyme treatment. The 400-kDa product is recognized by antibodies against the glycosaminoglycan-alpha domain and against synthetic NH2- and COOH-terminal peptides. Our preparations contain no major proteolytic products of versican, e.g. hyaluronectin or glial hyaluronate-binding protein. Having biochemical quantities of versican V2 available will allow us to test its putative modulatory role in neuronal cell adhesion and axonal growth.
Schmalfeldt
M
M
Institute of Clinical Pathology, Department of Pathology, University of Zürich, 8091 Zürich, Switzerland.
Dours-Zimmermann
M T
MT
Winterhalter
K H
KH
Zimmermann
D R
DR
D017398:Alternative Splicing; D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000378:metabolism; D001923:Brain Chemistry; D002417:Cattle; D011508:Chondroitin Sulfate Proteoglycans / Q000737:chemistry* / Q000235:genetics; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D016326:Extracellular Matrix Proteins / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry*; D037181:Lectins, C-Type; D008969:Molecular Sequence Data; D011509:Proteoglycans / Q000737:chemistry*; D011994:Recombinant Proteins / Q000276:immunology; D053675:Versicans
D000911:Antibodies, Monoclonal; D011508:Chondroitin Sulfate Proteoglycans; D018076:DNA, Complementary; D016326:Extracellular Matrix Proteins; D037102:Lectins; D037181:Lectins, C-Type; D011509:Proteoglycans; D011994:Recombinant Proteins; D053675:Versicans