Ultrafast Processes In Proteins And Other Assemblies

描述(由申请人提供)：通过跨膜(TM)螺旋、质子通道、快速折叠蛋白质、逆转录酶抑制剂和纤维中的相关水，将获得蛋白质结构随时间变化的残留量可视化，这是一种强大的新方法。膜蛋白对细胞生理学至关重要：它们包括细胞表面受体、离子通道、转运体和氧化还原蛋白。完整的膜蛋白介导细胞和细胞外基质之间的双向通讯，构成高等生物体所有编码序列的四分之一，超过一半的商业药物针对它们。尽管对了解人类健康至关重要，但对它们的3D结构和动力学的了解是有限的。具有双红外频率的2D IR将测量TM螺旋线波动的空间相关性。多肽和蛋白质以及氰基探针的同位素标记将提高2D IR的空间分辨率，并将其扩展到更大的生物组装。将研究GP A TM部分界面上的弱氢键，以显示键的运动以及它们如何稳定螺旋二聚体。2D IR揭示了膜上的空间排列、疏水效应、极性、氢键等埋藏残基之间的弱相互作用，揭示了螺旋缔合的机制和结构基础。这项研究从与病毒感染及其治疗、细胞信号和离子动员有关的化学键尺度动力学知识中寻求对生物组装中关键快速过程的分子水平的描述。水稳定和优化生命系统的动力学和功能，分子水平上对水与蛋白质和肽相互作用的动力学的描述是了解许多细胞过程的关键。甲型流感病毒M2原型质子通道的微观作用及其突变体如何逃脱抑制，将通过2D IR确定，从而有助于针对耐药形式的M2的抑制剂的设计。逆转录酶的研究将获得HIV酶抑制物复合体的分子水平知识。淀粉样纤维在阿尔茨海默病患者的脑组织中以斑块的形式积累，我们的2D IR实验涉及这些纤维中来自40个残基的多肽A？40的关键水通道。 公共卫生相关性：这项工作提供了流感病毒感染和治疗、蛋白质组装和信号转导、细胞表面受体和离子通道等方面的知识。用于HIV治疗的酶抑制剂和与阿尔茨海默病相关的淀粉样纤维的分子水平知识构成了这项工作的另一个重点。

DESCRIPTION (provided by applicant): A residue level visualization of protein structures changing with time through associated water in transmembrane (TM) helices, proton channels, fast folding proteins, reverse transcriptase inhibitors and fibrils will be obtained by two dimensional infrared spectroscopy (2D IR) a powerful new method. Membrane proteins are vital to cell physiology: they include cell-surface receptors, ion channels, transporters and redox proteins. Integral membrane proteins mediate bidirectional communication between cells and the extra cellular matrix, compose one-quarter of all coding sequences in higher organisms, and more than half of all commercial drugs target them. Though essential to understanding human health, knowledge of their 3D structures and their dynamics is limited. The 2D IR with dual IR frequencies, will measure spatial correlations in the fluctuations in TM helices. Isotopic labeling of peptides and proteins and nitrile probes will enhance the spatial resolution of 2D IR and extend it to larger biological assemblies. Weak hydrogen bonds at the interfaces of TM sections of Gp A will be examined to show the bond motions and how they stabilize helix dimers. 2D IR exposes spatial arrangements across the membrane, hydrophobic effects, polarity, hydrogen bonding and other weak interactions between buried residues that enlighten the mechanisms and structural basis of helix association. The research seeks a molecular level description of key fast processes in biological assemblies from a chemical bond scale dynamics knowledge pertaining to viral infections and their therapy, cellular signaling and ion mobilization. Water stabilizes and optimizes the dynamics and functionality of living systems and a molecular level description of the dynamics of water interacting with proteins and peptides is key to understanding many cellular processes The microscopic action of a prototype M2 proton channel from Influenza A virus and how its mutants can escape inhibition, will be determined by 2D IR thereby contributing to the design of inhibitors targeting drug-resistant forms of M2. The reverse transcriptase research will acquire molecular level knowledge of HIV enzyme-inhibitor complexes. Amyloid fibrils accumulate as plaques in the brain tissue of Alzheimer's patients and our 2D IR experiments concern key water channels in these fibrils from the 40-residue peptide A¿40. PUBLIC HEALTH RELEVANCE: The work provides a bond scale knowledge of influenza viral infections and therapy, protein assembly and signal transduction, cell-surface receptors and ion channels. A molecular level knowledge of enzyme-inhibitors for HIV treatments and amyloid fibrils associated with Alzheimer's disease form another focus of this work.