Protein Design Using Unnatural Amino Acids

描述(申请人提供)：膜蛋白约占人类蛋白质组的三分之一，在药物靶标中的比例过高。该项目的长期目标是能够明确和控制膜可溶性蛋白质组分的结构，以便它们可以用于治疗干预。可溶性蛋白质呈两部分结构，表面亲水，内部疏水。我们介绍了一种新的用于膜环境的二元图案化方案，该方案使用了高度氟化的氨基酸。本文建议的工作将集中在理解模型体系中含氟氨基酸相互作用的基本能量学，然后在水溶液中进行详细的表征。氟化氨基酸还将用于调节抗菌肽和治疗性多肽的生物活性和稳定性。此外，设计的结合在胶束和磷脂囊泡中的跨膜多肽将通过二硫键交换实验进行热力学表征，并测试它们在选择结合伙伴时的特异性。从上述研究中收集的信息将被用来在膜内创建离子通道和形成孔束。这些组件将使用生物物理技术进行进一步研究。这里提出的设计和表征研究应该促进膜转运剂的构建，并允许激活(或)抑制与治疗相关的膜包埋蛋白。

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.