Basement membrane collagen IV: Isolation of functional domains.

Basement membrane collagen IV: Isolation of functional domains.
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DOI:
10.1016/bs.mcb.2017.08.010
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发表时间:
2018
影响因子:
--
通讯作者:
Pedchenko VK
Pedchenko VK
中科院分区:
生物学4区
文献类型:
--
作者:
Boudko SP;Danylevych N;Hudson BG;Pedchenko VK

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IV 型胶原蛋白是基底膜的主要成分,基底膜是细胞外基质的特殊形式,为组织提供机械支撑,作为细胞粘附受体的多价配体和其他蛋白质的支架,并在组织发生、分化、体内平衡和重塑中发挥关键作用。 IV 型胶原蛋白是多种人类疾病发病机制的基础,包括古德帕斯彻氏病、阿尔波特综合征、糖尿病肾病、血管病和孔脑畸形。虽然从组织中分离 IV 型胶原蛋白分子是结构和功能研究的最终先决条件,但由于广泛的分子间交联以及与基底膜其他成分的非共价结合,它始终受到蛋白质不溶性的阻碍。在本章中,我们介绍了从基底膜或培养细胞沉积的细胞外基质中分离 IV 型胶原蛋白片段的方法,以及重组表达替代方案。这些方法有助于解决有关正常和病理条件下 IV 型胶原蛋白在组织发生中的作用的基本问题。
Collagen IV is a major constituent of basement membranes, specialized form of extracellular matrix that provides a mechanical support for tissues, serves as a polyvalent ligand for cell adhesion receptors and as a scaffold for other proteins, and plays a key role in tissue genesis, differentiation, homeostasis, and remodeling. Collagen IV underlies the pathogenesis of several human disorders including Goodpasture’s disease, Alport’s syndrome, diabetic nephropathy, angiopathy, and porencephaly. While the isolation of the collagen IV molecules from tissues is an ultimate prerequisite for structural and functional studies, it has been always hampered by the protein insolubility due to extensive intermolecular crosslinking and noncovalent associations with other components of basement membranes. In this chapter, we present methods for the isolation of collagen IV fragments from basement membranes or from extracellular matrix deposited by cultured cells, and the recombinant expression alternative. These methods are useful to address the fundamental questions on the role of collagen IV in tissue genesis under the normal and pathological conditions.
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