Rühland Claus, Schönherr Elke, Robenek Horst, Hansen Uwe, Iozzo Renato V, Bruckner Peter, Seidler Daniela G
Department of Physiological Chemistry and Pathobiochemistry, University of Münster, Germany.
FEBS J. 2007 Aug;274(16):4246-55. doi: 10.1111/j.1742-4658.2007.05951.x. Epub 2007 Jul 25.
Decorin is a multifunctional small leucine-rich proteoglycan involved in the regulation of collagen fibrillogenesis. In patients with a variant of Ehlers-Danlos syndrome, about half of the secreted decorin lacks the single glycosaminoglycan side chain. Notably, these patients have a skin-fragility phenotype that resembles that of decorin null mice. In this study, we investigated the role of glycanated and unglycanated decorin on collagen fibrillogenesis. Glycosaminoglycan-free decorin, generated by mutating Ser4 of the mature protein core into Ala (DCN-S4A), showed reduced inhibition of fibrillogenesis compared with the decorin proteoglycan. Interestingly, using a 3D matrix generated by decorin-null fibroblasts, an increase in fibril diameter was found after the addition of decorin, and even greater effects were observed with DCN-S4A. To avoid potential side effects of artificial tags, adenoviruses containing decorin and DCN-S4A were used to transduce decorin-null fibroblasts prior to matrix formation. Both molecules were efficiently incorporated into the matrix, with no changes in collagen composition and network formation, or altered expression of the related proteoglycan biglycan. Both decorin and DCN-S4A mutants increased the collagen fibril diameter, with the latter showing the most prominent effects. These data show that at early stages of fibrillogenesis, the glycosaminoglycan chain of decorin has a reducing effect on collagen fibril diameter.
核心蛋白聚糖是一种多功能的富含亮氨酸的小分子蛋白聚糖,参与胶原蛋白原纤维形成的调控。在患有埃勒斯-当洛综合征变异型的患者中,约一半分泌的核心蛋白聚糖缺乏单个糖胺聚糖侧链。值得注意的是,这些患者具有与核心蛋白聚糖基因敲除小鼠相似的皮肤脆弱表型。在本研究中,我们研究了糖基化和非糖基化核心蛋白聚糖在胶原蛋白原纤维形成中的作用。通过将成熟蛋白核心的Ser4突变为Ala(DCN-S4A)产生的无糖胺聚糖核心蛋白聚糖,与核心蛋白聚糖蛋白聚糖相比,对原纤维形成的抑制作用降低。有趣的是,使用由核心蛋白聚糖基因敲除的成纤维细胞产生的三维基质,添加核心蛋白聚糖后发现原纤维直径增加,而DCN-S4A的作用更明显。为避免人工标签的潜在副作用,在基质形成之前,使用含有核心蛋白聚糖和DCN-S4A的腺病毒转导核心蛋白聚糖基因敲除的成纤维细胞。两种分子都能有效地整合到基质中,胶原蛋白组成和网络形成没有变化,相关蛋白聚糖双糖链蛋白聚糖的表达也没有改变。核心蛋白聚糖和DCN-S4A突变体都增加了胶原原纤维直径,后者的作用最为显著。这些数据表明,在原纤维形成的早期阶段,核心蛋白聚糖的糖胺聚糖链对胶原原纤维直径有减小作用。