Patel Trushar R, Bernards Claudia, Meier Markus, McEleney Kevin, Winzor Donald J, Koch Manuel, Stetefeld Jörg
Department of Chemistry, University of Manitoba, 144 Dysart Road, Winnipeg, Manitoba RT3 2N2, Canada.
Institute for Dental Research and Oral Musculoskeletal Biology, Center for Molecular Medicine Cologne, Center for Biochemistry, Medical Faculty, University of Cologne, D-50931 Cologne, Germany.
Matrix Biol. 2014 Jan;33:60-7. doi: 10.1016/j.matbio.2013.07.009. Epub 2013 Aug 12.
Nidogen-1 is a key basement membrane protein that is required for many biological activities. It is one of the central elements in organizing basal laminae including those in the skin, muscle, and the nervous system. The self-assembling extracellular matrix that also incorporates fibulins, fibronectin and integrins is clamped together by networks formed between nidogen, perlecan, laminin and collagen IV. To date, the full-length version of nidogen-1 has not been studied in detail in terms of its solution conformation and shape because of its susceptibility to proteolysis. In the current study, we have expressed and purified full-length nidogen-1 and have investigated its solution behavior using size-exclusion chromatography (SEC), dynamic light scattering (DLS) and small angle X-ray scattering (SAXS). The ab initio shape reconstruction of the complex between nidogen-1 and the laminin γ-1 short arm confirms that the interaction is mediated solely by the C-terminal domains: the rest of the domains of both proteins do not participate in complex formation.
巢蛋白-1是一种关键的基底膜蛋白，许多生物活动都需要它。它是组织基底层（包括皮肤、肌肉和神经系统中的基底层）的核心成分之一。由巢蛋白、基底膜聚糖、层粘连蛋白和IV型胶原之间形成的网络，将还包含纤维蛋白、纤连蛋白和整合素的自组装细胞外基质紧密结合在一起。迄今为止，由于巢蛋白-1易被蛋白水解，其全长版本的溶液构象和形状尚未得到详细研究。在本研究中，我们表达并纯化了全长巢蛋白-1，并使用尺寸排阻色谱法（SEC）、动态光散射（DLS）和小角X射线散射（SAXS）研究了其溶液行为。巢蛋白-1与层粘连蛋白γ-1短臂之间复合物的从头形状重建证实，这种相互作用仅由C端结构域介导：两种蛋白质的其余结构域不参与复合物的形成。
