Solid-state Nanopore Identification for Proteins

描述（由申请人提供）：这项拟议的研究是通过小孔（氮化硅膜中的固态纳米孔）吸引变性蛋白质的单分子，该小孔是灵敏检测器不可或缺的一部分。固态纳米孔检测器被设计为保证在电场的驱动下，每个多肽以连续的单行顺序穿过纳米孔。每个多肽的移位将诱导瞬时电子信号，即检测器中的电流阻断。研究目标是开发纳米孔技术，通过测量电流封锁来记录单个多肽的易位，探测肽的基本特性，包括它们的长度，直径，二级结构，电荷，并最终以高速，高分辨率和低成本的氨基酸序列。具体目标是：1.在离子水溶液中，开发由电场驱动的多肽链通过固态纳米孔移位的单通道记录。这种记录将用于实现多肽链基本性质的电子读出。研究电子信号如何与这些感兴趣的基本分子特性相关。2.开发对固态纳米孔的厚度、化学活性、导电性和噪声特性的可靠控制，以符合单一蛋白质传感的要求。3.研究并测试了合适的温度、pH、偏压和纳米孔尺寸等条件，优化了固态纳米孔传感器对氨基酸链移位和识别的过程。4.开发数据分析软件和统计模型来解释纳米孔电子信号。如果这里提出的研究目标得以实现，一个高通量的设备，可以探测和直接“读取”电子，在单分子水平上，大小，电荷，折叠，和蛋白质的序列，将大大改变生物学和医学科学的步伐。单肽链的分子和原子特征的高通量纳米孔探针的开发，以及使这种探针的开发成为可能的基本理解，将彻底改变功能基因组学和蛋白质组学。

DESCRIPTION (provided by applicant): This proposed research is to draw single molecules of denatured proteins through a small pore, a solid state nanopore in silicon nitride membrane, that is integral to a sensitive detector. The solid state nanopore detector is designed to guarantee that driven by electric field, each polypeptide traverses the nanopore in sequencial, single file order. The translocation of each polypeptide will induce a transient electronic signal, a current blockade in the detector. The research goal is to develop the nanopore technique to record single polypeptide translocations by measuring the current blockades, probe the peptide's fundamental properties including their length, diameter, secondary structure, charge, and eventually the amino acid sequence at high speed, high resolution, and low cost. The specific aims are: 1. Develop single channel recordings of polypeptide chains translocating through solid state nanopores driven by electric field in aqueous ionic solution. Such recordings will be used to achieve an electronic read-out of the fundamental properties of polypeptide chains. Study how the electronic signals are related to these fundamental molecular properties of interest. 2. Develop reliable control over the thickness, chemical activity, electrical conduction and noise properties of solid state nanopores compatible with the requirements of single protein sensing. 3. Study and test the appropriate temperature, pH, bias voltage and nanopore size conditions for optimize the process of amino acid chain translocation and identification through solid state nanopore sensors. 4. Develop data analysis software and statistical models to interpret nanopore electronic signals. If the research goals proposed here are reached, a high-throughput device that can probe and directly "read" electronically, at the single molecule level, the size, charge, folding, and sequence of proteins, will dramatically alter the pace of biology and medical science. The development of high throughput nanopore probes of the molecular and atomic characteristics of single peptide chains, and the fundamental understanding that will make development of such probes possible, would revolutionize functional genomics, and proteomics.