AMINO ACID/PEPTIDE TRANSFER THERMODYNAMICS

我们两个小组将合作开发色谱方法， 测量氨基酸分配，以更好地了解蛋白质 稳定性 目前的实验涉及液体油/水分配 或结晶二酮哌嗪溶解。但蛋白质核心既不是 不像液体油也不像晶体二肽。 我们建议3 色谱方法：（1）有序油水分配 色谱，在其中我们将有独立的控制空间位阻 限制和油中的“液-固”程度。 （二） 反相氢键色谱法，我们将有 独立控制油和水溶液中的氢键能力 阶段。(3)肽相色谱法，我们将共价 将肽而不是烷基链连接到二氧化硅载体上， 从水性溶剂中分离氨基酸。 我们将测量 传递自由能、熵、熵和热容， 氨基酸和肽。我们的目标是：（一）开发分区 比目前的实验更接近蛋白质核心的介质 模型，和（ii）解剖氢键和空间组分， 传递热力学。 A（4）理论部分将尝试 用这些和其他实验来解释蛋白质的稳定性 一个理论的努力将使用快速搜索策略（“疏水拉链”） 结合分配数据来模拟蛋白质稳定性。 我们 最终目标是利用这些氨基酸的分配研究， 肽来帮助解开蛋白质的驱动力 稳定性

Our two groups will collaborate to develop chromatography methods to measure amino acid partitioning, to better understand protein stabilities. Current experiments involve liquid oil/water partitioning or crystal diketopiperazine dissolution. But protein cores are neither like liquid oils nor like crystalline dipeptides. We propose 3 chromatographic methods: (1) Ordered-Oil/Water Partitioning Chromatography, in which we will have independent control over the steric constraints and the degree of "liquidness-solidness" in the oil. (2) Reversed-Phase Hydrogen Bonding Chromatography, in which we will have independent control over the hydrogen bonding capacity in oil and aqueous phases. (3) Peptide Phase Chromatography, in which we will covalently attach peptides, rather than alkyl chains, to silica supports for the partitioning of amino acids from aqueous solvents. We will measure transfer free energies, enthalpies, entropies, and heat capacities for amino acids and peptides. Our goals are: (i) to develop partitioning media that more closely resemble protein cores than current experimental models, and (ii) to dissect hydrogen-bonding and steric components from the transfer thermodynamics. A (4) Theoretical Component will attempt to interpret protein stabilities in terms of these and other experiments. A theory effort will use a fast search strategy ("hydrophobic Zipper") combined with the partitioning data to model protein stabilities. Our ultimate goal is to use these partitioning studies of amino acids and peptides to help unravel the driving forces underlying protein stabilities.