Werneck Claudio C, Trask Barbara Crippes, Broekelmann Thomas J, Trask Timothy M, Ritty Timothy M, Segade Fernando, Mecham Robert P
Department of Cell Biology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Biol Chem. 2004 May 28;279(22):23045-51. doi: 10.1074/jbc.M402656200. Epub 2004 Mar 24.
Using yeast two-hybrid, ligand blotting, and solid phase binding assays, we have shown that microfibril-associated glycoprotein-1 (MAGP-1) interacts with the 8-cysteine motif of fibrillin-2 encoded by exon 24. Binding to this sequence was demonstrated for full-length MAGP-1 as well as for the MAGP-1 matrix-binding domain encoded by exons 7 and 8. The matrix-binding domain, but not the full-length protein, also bound to regions of fibrillin-2 defined by exons 16 and 17, exon 20, and exons 23 and 24. Interestingly, no binding was detected to sequences near the N or C terminus where MAGP-1 and MAGP-2, respectively, were shown to interact with fibrillin-1. The localization of MAGP-1 binding to the 8-Cys domain encoded by exon 24 suggests that the bead structure of microfibrils consists of exon 24 and portions of the central region of fibrillin-2. Exon 24 in fibrillin lies in the region of the molecule where mutations produce the most severe phenotypes associated with Marfan syndrome (fibrillin-1) and congenital contractural arachnodactyly (fibrillin-2). It is possible that these mutations alter the ability of fibrillin to bind MAGP-1, which may contribute to the severity of the disease.
利用酵母双杂交、配体印迹和固相结合试验,我们已表明微原纤维相关糖蛋白-1(MAGP-1)与由外显子24编码的原纤蛋白-2的8个半胱氨酸基序相互作用。全长MAGP-1以及由外显子7和8编码的MAGP-1基质结合结构域均显示与该序列结合。基质结合结构域而非全长蛋白,也与由外显子16和17、外显子20以及外显子23和24所界定的原纤蛋白-2区域结合。有趣的是,未检测到与MAGP-1和MAGP-2分别被证明与原纤蛋白-1相互作用的N或C末端附近序列的结合。MAGP-1与由外显子24编码的8个半胱氨酸结构域的结合定位表明,微原纤维的珠状结构由外显子24和原纤蛋白-2中央区域的部分组成。原纤蛋白中的外显子24位于分子中突变产生与马凡综合征(原纤蛋白-1)和先天性挛缩性蜘蛛指症(原纤蛋白-2)相关的最严重表型的区域。有可能这些突变改变了原纤蛋白结合MAGP-1的能力,这可能导致疾病的严重程度。