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 |
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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 Ã
, a maximum diameter of 98 ± 5 Ã
, and approximates
two intertwined C shapes. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M211936200 |