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Structure, Function and Application of Plant Nonspecific Lipid Transfer Protein
Dissertation

Structure, Function and Application of Plant Nonspecific Lipid Transfer Protein

Chao-Sheng Cheng
Doctor of Philosophy (PHD), 國立清華大學, 生命科學系
2005

Abstract

植物非專一性脂質運輸蛋白 疏水性空洞 蛋白質穩定度 比較模擬法 嵌合 演化樹 植物抗菌 化學位移分布 氫/氘交換 藥物運輸 快速篩選法 Plant non-specific lipid transfer proteins Hydrophobic cavity Protein stability Comparative modeling Docking Phylogenetic tree Plant defense Chemical shift perturbations H/D exchange Drug carrier High throughput screening method
Plant nonspecific lipid transfer proteins (nsLTPs) are small basic proteins which are characterized by their ability to transfer lipids between membranes in vitro. Based on the molecular weight, nsLTPs are classified into two subfamilies, nsLTP1 (9 kDa) and nsLTP2 (7 kDa). In contrast with nsLTP1, only rice and wheat nsLTP2 structures have been determined in this subfamily. In order to realize nsLTP2s, comparative modeling and molecular docking were used to study their structural, biophysical and biochemical properties. Besides, nsLTPs seem to play an important role in plant defense although the exact mechanism for antimicrobial effects is still unclear. We purpose that nsLTPs may associate with a sterol molecule to trigger the plant defense. This hypothesis was verified by using various spectroscopic techniques like fluorescence, circular dichroism, mass spectrometry and NMR. We also used site-directed mutagenesis to investigate the influence of hydrophobic residues on sterol binding and protein stability. Finally, the wide binding properties allow nsLTPs to be a potential drug carrier. Therefore, we used a computer-based high throughput screening (HTS) method to screen drug library to identify the possible binding drugs, thus increasing the added value for nsLTPs in drug delivery systems.

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