Chimeric fibronectin matrix mimetic as a functional growth- and migration-promoting adhesive substrate.
Chimeric fibronectin matrix mimetic as a functional growth- and migration-promoting adhesive substrate.
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DOI：
10.1016/j.biomaterials.2010.11.050
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发表时间：
2011-03
期刊：
影响因子：
14
通讯作者：
Hocking, Denise C.
中科院分区：
文献类型：
--
作者：
Roy, Daniel C.;Wilke-Mounts, Susan J.;Hocking, Denise C.
治疗性蛋白质工程结合了遗传、生物化学和功能信息，以改善现有蛋白质或发明新的蛋白质技术。利用这些原理，我们开发了一种方法来提供细胞外基质（ECM）纤连蛋白特异性信号的细胞。纤连蛋白基质组装是将纤连蛋白的无活性可溶形式转化为生物活性ECM原纤维的细胞依赖性过程。ECM纤连蛋白刺激正常组织再生所需的细胞功能，包括细胞生长、扩散、迁移和胶原重组。我们已经开发了重组纤连蛋白片段，其通过将纤连蛋白的第一个III型重复序列（FNIII 1H）的隐蔽肝素结合片段偶联至整联蛋白结合结构域（FNIII 8 -10）来模拟ECM纤连蛋白对细胞功能的影响。GST/III 1H，8-10支持细胞粘附和铺展，并比血浆纤连蛋白更大程度地刺激细胞增殖。产生缺失和位点特异性突变体构建体以鉴定GST/III 1H，8-10中的活性区并减小构建体大小。一种嵌合构建体，其中整合素结合RGDS环插入FNIII 8（GST/III 1H，8 RGD）中的类似位点，支持细胞粘附和迁移，并增强细胞增殖和胶原凝胶收缩。GST/III 1H，8 RGD在细菌中表达，并通过亲和层析从可溶性裂解物级分中纯化。纤连蛋白基质组装通常响应于组织损伤而上调。ECM纤连蛋白水平的降低与不愈合的伤口相关。工程化的纤连蛋白基质模拟物绕过了慢性伤口中对细胞依赖性纤连蛋白基质组装的需要，是刺激对组织修复至关重要的细胞活性的新方法。
Therapeutic protein engineering combines genetic, biochemical, and functional information to improve existing proteins or invent new protein technologies. Using these principles, we developed an approach to deliver extracellular matrix (ECM) fibronectin-specific signals to cells. Fibronectin matrix assembly is a cell-dependent process that converts the inactive, soluble form of fibronectin into biologically-active ECM fibrils. ECM fibronectin stimulates cell functions required for normal tissue regeneration, including cell growth, spreading, migration, and collagen reorganization. We have developed recombinant fibronectin fragments that mimic the effects of ECM fibronectin on cell function by coupling the cryptic heparin-binding fragment of fibronectin’s first type III repeat (FNIII1H) to the integrin-binding domain (FNIII8-10). GST/III1H,8-10 supports cell adhesion and spreading and stimulates cell proliferation to a greater extent than plasma fibronectin. Deletion and site-specific mutant constructs were generated to identify the active regions in GST/III1H,8-10 and reduce construct size. A chimeric construct in which the integrin-binding, RGDS loop was inserted into the analogous site in FNIII8 (GST/III1H,8RGD), supported cell adhesion and migration, and enhanced cell proliferation and collagen gel contraction. GST/III1H,8RGD was expressed in bacteria and purified from soluble lysate fractions by affinity chromatography. Fibronectin matrix assembly is normally up-regulated in response to tissue injury. Decreased levels of ECM fibronectin are associated with non-healing wounds. Engineering fibronectin matrix mimetics that bypass the need for cell-dependent fibronectin matrix assembly in chronic wounds is a novel approach to stimulating cellular activities critical for tissue repair.
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DOI：
10.1083/jcb.95.2.369
发表时间：
1982-11
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hynes RO;Yamada KM
通讯作者：
Yamada KM
影响因子：
21.3
作者：
Balaban, NQ;Schwarz, US;Geiger, B
通讯作者：
Geiger, B
DOI：
10.1073/pnas.91.21.10114
发表时间：
1994-10-11
影响因子：
11.1
作者：
ERICKSON, HP
通讯作者：
ERICKSON, HP
影响因子：
4.8
作者：
CHUNG, CY;ZARDI, L;ERICKSON, HP
通讯作者：
ERICKSON, HP
DOI：
10.1083/jcb.200112031
发表时间：
2002-07-08
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hocking DC;Kowalski K
通讯作者：
Kowalski K