Solid-state Nanopore Identification for Proteins

描述（由申请人提供）：这项拟议的研究是通过一个小孔（氮化硅膜上的固态纳米孔）来绘制变性蛋白质的单分子，这是一个敏感探测器的组成部分。固体纳米孔探测器是为了保证在电场的驱动下，每个多肽按顺序、单排顺序穿过纳米孔而设计的。每一个多肽的易位都会引起一个瞬态电子信号，即检测器中的电流封锁。研究目标是开发纳米孔技术，通过测量当前的阻滞来记录单个多肽的易位，以高速、高分辨率和低成本探测肽的基本性质，包括它们的长度、直径、二级结构、电荷和最终的氨基酸序列。具体目标是：1。建立多肽链在电场驱动下在离子水溶液中通过固态纳米孔转移的单通道记录。这样的记录将被用来实现多肽链基本性质的电子读出。研究电子信号与这些感兴趣的基本分子性质之间的关系。2. 开发可靠的厚度控制，化学活性，电导率和噪声性能的固态纳米孔兼容的要求，以满足单蛋白传感。3. 研究和测试合适的温度、pH、偏置电压和纳米孔尺寸条件，以优化通过固态纳米孔传感器进行氨基酸链易位和鉴定的过程。4. 开发数据分析软件和统计模型来解释纳米孔电子信号。如果本文提出的研究目标得以实现，那么一种可以在单分子水平上探测并直接“读取”蛋白质的大小、电荷、折叠和序列的高通量设备将极大地改变生物学和医学科学的步伐。单肽链分子和原子特征的高通量纳米孔探针的开发，以及使此类探针开发成为可能的基本认识，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.