Fibrillin-1: Organization in microfibrils and structural properties
Fibrillin-1: Organization in microfibrils and structural properties
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DOI：
10.1006/jmbi.1996.0237
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发表时间：
1996-04-26
影响因子：
5.6
通讯作者：
Sakai, LY
中科院分区：
文献类型：
--
作者：
Reinhardt, DP;Keene, DR;Sakai, LY
为了研究纤维蛋白-1的微纤维组织和结构特性，我们在人类细胞中生产了重叠的重组多肽，它们总共跨越了纤维蛋白-1分子。在非变性条件下对多肽进行了纯化，广泛的表征表明折叠正确。纯化的蛋白用于标记单抗26、69和201。结合位点位于氨基酸残基45-450(MAb26)、451-909(MAb201)之间的N端，以及位于氨基酸残基2093-2871(MAb69)之间的C端。这些抗体对扩展的珠状结构(微纤丝)的免疫定位表明，纤维蛋白-1的N-末端和C-末端位于珠子附近和相对的两侧，分子的更中心部分位于珠子之间。每个表位在每个珠子之间出现一次。这些数据为微原纤维中纤维蛋白的组织提供两种可能的模型。然而，重组多肽上的抗体结合位置与组织中标记事件之间的距离比较表明，纤维蛋白分子在其组织形式中以微纤维的形式致密。对重组多肽的进一步分析提供了关于8-半胱氨酸基序的新信息，8-半胱氨酸基序是存在于纤维蛋白和TGFP结合蛋白中的一个新结构域，并表明纤维蛋白在其N-末端和C-末端被加工。(C)1996年学术出版社有限公司
To investigate the microfibrillar organization and structural properties of fibrillin-1, we produced overlapping recombinant peptides in human cells which altogether span the fibrillin-1 molecule. The peptides were purified under non-denaturing conditions and extensive characterization indicated correct folding. The purified proteins were used to map monoclonal antibodies 26, 69 and 201. The binding sites are located at the N-terminal end between amino acid residues 45 and 450 (mAb 26), 451 and 909 (mAb 201) and at the C-terminal end between residues 2093 and 2871 (mAb 69). Immunolocalization of these antibodies to extended beaded structures (microfibrils) demonstrated that the N- and C-terminal ends of fibrillin-1 are located in proximity and on opposite sides of the beads, and more central parts of the molecule are located between the beads. Each epitope is present once between each bead. These data allow two possible models for the organization of fibrillin in microfibrils. However, comparison of distances between antibody binding sites on the recombinant peptides and labeling events in tissue suggests that fibrillin molecules are compacted within their tissue form as microfibrils. Additional analyses of the recombinant peptides provide new information regarding the eight-cysteine motif, a novel domain present in fibirillins and TGFP binding proteins, and suggest that fibrillins are processed at their N- and C-terminal ends. (C) 1996 Academic Press Limited