Versican V2 Is a Major Extracellular Matrix Component of the Mature Bovine Brain*
- Michael Schmalfeldt‡,
- Marı́a T. Dours-Zimmermann‡,
- Kaspar H. Winterhalter§ and
- Dieter R. Zimmermann‡¶
- From the ‡Institute of Clinical Pathology, Department of Pathology, University of Zürich, 8091 Zürich, and the §Laboratory of Biochemistry I, Federal Institute of Technology, 8092 Zürich, Switzerland
Abstract
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-α 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.
Footnotes
-
↵* This work was supported by grants from the Krebsliga des Kantons Zürich and from the Lydia Hochstrasser Foundation (to D. R. Z.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) AF060456, AF060457, AF060458, and AF060459 for versican V0, V1, V2, and V3, respectively.
-
↵¶ To whom correspondence should be addressed: Dept. of Pathology, Institute of Clinical Pathology, University of Zürich, Schmelzbergstrasse 12, CH-8091 Zürich, Switzerland. Tel.: 41-1-255-3945 Fax: 41-1-255-4508 E-mail:dieterzi{at}pathol.unizh.ch.
-
↵1 The abbreviations used are: GAG, glycosaminoglycan; PCR, polymerase chain reaction; PAGE, polyacrylamide gel electrophoresis; kb, kilobase(s).
-
- Received February 3, 1998.
- Revision received April 8, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
