Light and X-ray Scattering Show Decorin to Be a Dimer in Solution

Decorin is a widely distributed member of the extracellular matrix small leucine-rich repeat glycoprotein/proteoglycan family. For investigation of its physical properties, decorin from two sources (young steer skin and a recombinant adenovirus) was used. The first sample was extracted into 7 m urea...

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Veröffentlicht in:The Journal of biological chemistry 2003-05, Vol.278 (20), p.18353-18359
Hauptverfasser: Scott, Paul G, Grossmann, J Gunter, Dodd, Carole M, Sheehan, John K, Bishop, Paul N
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Sprache:eng
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Zusammenfassung:Decorin is a widely distributed member of the extracellular matrix small leucine-rich repeat glycoprotein/proteoglycan family. For investigation of its physical properties, decorin from two sources (young steer skin and a recombinant adenovirus) was used. The first sample was extracted into 7 m urea and purified, while the second was isolated from medium conditioned by 293A cells infected with adenovirus and purified without chaotropes. The only chemical differences detected between these materials were a slightly shorter glycosaminoglycan chain and the retention of the propeptide on the latter. Circular dichroism spectra of the two samples were virtually identical, showing a high proportion of β-sheet and β-turn and little α-helix. The protein cores were completely denatured in 2.25 m guanidine HCl (GdnHCl) but recovered their secondary structure on removal of chaotrope. Light scattering of material eluted from gel-filtration columns in Tris-buffered saline, pH 7.0, gave molecular mass values of 165 ± 1 kDa and 84.6 ± 4 kDa for intact decorin and the glycoprotein core produced by digestion with chondroitin ABC lyase, respectively. Intact recombinant prodecorin had a mass of 148 ± 18 kDa. These values, which are double those estimated from SDS gel electrophoresis or from the known sequences and compositions, were halved in 2.5 m GdnHCl. Data from solution x-ray scattering of intact decorin and its core in Tris-buffered saline are consistent with a dimeric particle whose protein component has a radius of gyration of 31.6 ± 0.4 Å, a maximum diameter of 98 ± 5 Å, and approximates two intertwined C shapes.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M211936200