Protein Design Using Unnatural Amino Acids

描述（由申请人提供）：膜蛋白约占人类蛋白质组的三分之一，并且在药物靶标中占比过高。这个项目的长期目标是能够指定和控制膜可溶性蛋白成分的结构，以便它们可以用于治疗干预。可溶性蛋白呈两部分结构，外部亲水，内部疏水。我们介绍了一种新的二元模式方案，用于膜环境中使用高氟化氨基酸。这里提出的工作将集中在理解模型系统中氟化氨基酸相互作用的基本能量学，然后在水溶液中进行详细表征。氟化氨基酸也将用于调节抗菌和治疗肽的生物活性和稳定性。此外，设计的在胶束和磷脂囊泡中结合的跨膜肽将使用二硫交换法进行热力学表征，并测试其选择结合伙伴的特异性。从上述研究中收集的信息将用于在膜内创建离子通道和孔形成束。这些组合将使用生物物理技术进一步研究。本文提出的设计和表征研究应促进膜转运剂的构建，并允许激活（或）抑制治疗相关的膜嵌入蛋白。

DESCRIPTION (provided by applicant): Membrane proteins make up roughly a third of the human proteome and are over represented in drug targets. The broad long-term goal of this project is to be able to specify and control structure of membrane soluble protein components so that they can be used in therapeutic intervention. Soluble proteins display a bipartite architecture with hydrophilic exteriors and hydrophobic interiors. We have introduced a novel binary patterning scheme for use in membrane environments by use of highly fluorinated amino acids. The work proposed here will focus on understanding fundamental energetics of the interaction of fluorinated amino acids in model systems followed by detailed characterization in aqueous solution. Fluorinated amino acids will also be used to modulate the bioactivity and stability of antimicrobial and therapeutic peptides. Furthermore, designed transmembrane peptides that associate in micelles and phospholipid vesicles will be thermodynamically characterized using disulfide exchange assays and tested for their specificity in choosing binding partners. The information gleaned from the above studies will be used to create ion channel and pore forming bundles within membranes. These assemblies will be further studied using biophysical techniques. The design and characterization studies proposed here should facilitate the construction of membrane transport agents and also allow for activation (or) inhibition of therapeutically relevant membrane embedded proteins.