Van den Bergh Françoise, Fu Chang-Ling, Olague-Marchan Monica, Giudice George J
Department of Dermatology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA.
Biochem Biophys Res Commun. 2006 Dec 1;350(4):1032-7. doi: 10.1016/j.bbrc.2006.09.147. Epub 2006 Oct 5.
Collagen XVII/BP180 is a transmembrane constituent of the epidermal anchoring complex. To study the role of its non-collagenous linker domain, NC16A, in protein assembly and stability, we analyzed the following recombinant proteins: the collagen XVII extracellular domain with or without NC16A, and a pair of truncated proteins comprising the COL15-NC15 stretch expressed with or without NC16A. All four proteins were found to exist as stable collagen triple helices; however, the two missing NC16A exhibited melting temperatures significantly lower than their NC16A-containing counterparts. Protein refolding experiments revealed that the rate of triple helix assembly of the collagen model peptide GPP(10) is greatly increased by the addition of an upstream NC16A domain. In summary, the NC16A linker domain of collagen XVII exhibits a positive effect on both the rate of assembly and the stability of the adjoining collagen structure.
胶原蛋白 XVII/BP180 是表皮锚定复合体的一种跨膜成分。为了研究其非胶原蛋白连接域 NC16A 在蛋白质组装和稳定性中的作用，我们分析了以下重组蛋白：带有或不带有 NC16A 的胶原蛋白 XVII 细胞外结构域，以及一对包含 COL15-NC15 片段且带有或不带有 NC16A 表达的截短蛋白。发现所有这四种蛋白均以稳定的胶原三螺旋形式存在；然而，缺少 NC16A 的两种蛋白的解链温度明显低于其含 NC16A 的对应物。蛋白质复性实验表明，通过添加上游 NC16A 结构域，胶原模型肽 GPP(10) 的三螺旋组装速率大大提高。总之，胶原蛋白 XVII 的 NC16A 连接域对组装速率和相邻胶原结构的稳定性均具有积极作用。
