Gelatin Binding to the 6F11F22F2 Fragment of Fibronectin Is Independent of Module-Module Interactions

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• 作者：Amanda Pagett ; Iain D. Campbell ; and Andrew R. Pickford
• 刊名：Biochemistry
• 出版年：2005
• 出版时间：November 8, 2005
• 年：2005
• 卷：44
• 期：44
• 页码：14682 - 14687
• 全文大小：348K
• 年卷期：v.44,no.44(November 8, 2005)
• ISSN：1520-4995

文摘

Fibronectin, a large modular protein, interacts with many other proteins in the extracellularmatrix and on the cell surface. It has previously been shown that interactions between noncontiguousmodules exist in the collagen binding region. It is shown here that the interaction between the sixth typeI module (⁶F1) and the second type II module (²F2) can be disrupted by mutation of a residue in theintermodule interface of the ⁶F1¹F2²F2 fragment. The perturbation of the interface and the binding ofcollagen-derived peptides to individual modules were assessed by high-resolution nuclear magneticresonance (NMR) spectroscopy. Cooperativity between the modules in binding ligand was assessed byanalytical gelatin affinity chromatography of the mutant and wild-type proteins. Differential scanningcalorimetry (DSC) was used to probe the influence of the interface on module stability. It is shown thatwhile the ⁶F1-²F2 interface confers significant thermal stability to the ²F2 module, it has little effect ongelatin binding activity of the ⁶F1¹F2²F2 fragment.