Solid-state Nanopore Identification for Proteins

描述(申请人提供)：这项拟议的研究是通过一个小孔提取变性蛋白质的单分子，该小孔是氮化硅薄膜中的一个固态纳米孔，是灵敏探测器的组成部分。固态纳米孔探测器的设计是为了保证在电场的驱动下，每个多肽按顺序、单列顺序遍历纳米孔。每一个多肽的移位都会诱导一个瞬间的电子信号，也就是探测器中的电流阻塞。研究目标是开发纳米孔技术，通过测量当前的堵塞来记录单个多肽的易位，探测多肽的基本性质，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方式获得氨基酸序列。其具体目标是：1.发展离子水溶液中电场驱动下多肽链通过固态纳米孔转移的单通道记录。这种记录将被用来实现多肽链的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属性相关。2.开发符合单个蛋白质传感要求的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。3.对固态纳米孔传感器优化氨基酸链转移和识别的适宜温度、pH、偏压和纳米孔尺寸条件进行了研究和试验。4.开发数据分析软件和统计模型，解释纳米孔电信号。如果达到这里提出的研究目标，一种高通量设备将极大地改变生物学和医学科学的步伐，这种设备可以在单分子水平上用电子手段探测和直接读取蛋白质的大小、电荷、折叠和序列。单肽链分子和原子特征的高通量纳米孔探针的开发，以及使此类探针的开发成为可能的基本认识，将使功能基因组学和蛋白质组学发生革命性的变化。

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.