公开(公告)号：US08669116B2

公开(公告)日：2014-03-11

申请号：US10506877

申请日：2003-03-11

申请人： Steven P. Gygi ,  Junmin Peng

发明人： Steven P. Gygi ,  Junmin Peng

IPC分类号： C12Q1/37 ,  C07K1/22 ,  G01N33/68

CPC分类号： G01N33/6848 ,  C07K1/22 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/6803 ,  G01N33/6842 ,  G01N33/6896 ,  G01N2440/36 ,  G01N2560/00 ,  Y10T436/24

摘要： The invention provides a method detecting and quantifying proteins by mass spectrophotometric analysis using peptide internal standards and provides a highly sensitive way of detecting protein modifications. In one aspect, the invention provides a method for determining a site of ubiquitination in a polypeptide and for evaluating ubiquitination targets in a population of polypeptides. In this way, a proteome ubiquitination map can be obtained which comprises information relating to the ubiquitination states of a plurality of cellular polypeptides. Maps can be obtained for a variety of different types of cells and cell states. For example, ubiquitination targets in normal and diseased cells can be evaluated. Preferably, the map is stored as data files in a database. Individual ubiquitinated polypeptides identified can be used to generate molecular probes diagnostic of a cell state and/or can serve as targets for agents that modulate one or more cellular processes.

摘要翻译： 本发明提供了通过使用肽内标的质谱分析法检测和定量蛋白质的方法，并提供了检测蛋白质修饰的高度灵敏的方法。 一方面，本发明提供了一种确定多肽泛素化位点的方法，并用于评价多肽群体中的泛素化靶标。 以这种方式，可以获得包含与多个细胞多肽的泛素化状态有关的信息的蛋白质组泛素化图。 可以获得各种不同类型的细胞和细胞状态的图。 例如，可以评估正常和患病细胞中的泛素化靶标。 优选地，地图作为数据文件存储在数据库中。 鉴定的单个泛素化多肽可用于产生诊断细胞状态的分子探针和/或可用作调节一种或多种细胞过程的试剂的靶标。

公开(公告)号：US20060148093A1

公开(公告)日：2006-07-06

申请号：US10506877

申请日：2003-03-11

申请人： Steven Gygi ,  Junmin Peng

发明人： Steven Gygi ,  Junmin Peng

IPC分类号： G01N24/00

CPC分类号： G01N33/6848 ,  C07K1/22 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/6803 ,  G01N33/6842 ,  G01N33/6896 ,  G01N2440/36 ,  G01N2560/00 ,  Y10T436/24

摘要： The invention provides a method detecting and quantifying proteins by mass spectrophotometric analysis using peptide internal standards and provides a highly sensitive way of detecting protein modifications. In one aspect, the invention provides a method for determining a site of ubiquitination in a polypeptide and for evaluating ubiquitination targets in a population of polypeptides. In this way, a proteome ubiquitination map can be obtained which comprises information relating to the ubiquitination states of a plurality of cellular polypeptides. Maps can be obtained for a variety of different types of cells and cell states. For example, ubiquitination targets in normal and diseased cells can be evaluated. Preferably, the map is stored as data files in a database. Individual ubiquitinated polypeptides identified can be used to generate molecular probes diagnostic of a cell state and/or can serve as targets for agents that modulate one or more cellular processes.

摘要翻译： 本发明提供了通过使用肽内标的质谱分析法检测和定量蛋白质的方法，并提供了检测蛋白质修饰的高度灵敏的方法。 一方面，本发明提供了一种确定多肽泛素化位点的方法，并用于评价多肽群体中的泛素化靶标。 以这种方式，可以获得包含与多个细胞多肽的泛素化状态有关的信息的蛋白质组泛素化图。 可以获得各种不同类型的细胞和细胞状态的图。 例如，可以评估正常和患病细胞中的泛素化靶标。 优选地，地图作为数据文件存储在数据库中。 鉴定的单个泛素化多肽可用于产生诊断细胞状态的分子探针和/或可用作调节一种或多种细胞过程的试剂的靶标。

公开(公告)号：US20120149883A1

公开(公告)日：2012-06-14

申请号：US13284255

申请日：2011-10-28

申请人： Steven P. Gygi ,  Junmin Peng

发明人： Steven P. Gygi ,  Junmin Peng

IPC分类号： C07K17/00 ,  C07K16/00

CPC分类号： G01N33/6848 ,  C07K1/22 ,  C07K7/08 ,  C12Q1/37 ,  G01N33/6803 ,  G01N33/6842 ,  G01N33/6896 ,  G01N2440/36 ,  G01N2560/00 ,  Y10T436/24

摘要： The invention provides a method detecting and quantifying proteins by mass spectrophotometric analysis using peptide internal standards and provides a highly sensitive way of detecting protein modifications. In one aspect, the invention provides a method for determining a site of ubiquitination in a polypeptide and for evaluating ubiquitination targets in a population of polypeptides. In this way, a proteome ubiquitination map can be obtained which comprises information relating to the ubiquitination states of a plurality of cellular polypeptides. Maps can be obtained for a variety of different types of cells and cell states. For example, ubiquitination targets in normal and diseased cells can be evaluated. Preferably, the map is stored as data files in a database. Individual ubiquitinated polypeptides identified can be used to generate molecular probes diagnostic of a cell state and/or can serve as targets for agents that modulate one or more cellular processes.

摘要翻译： 本发明提供了通过使用肽内标的质谱分析法检测和定量蛋白质的方法，并提供了检测蛋白质修饰的高度灵敏的方法。 一方面，本发明提供了一种确定多肽泛素化位点的方法，并用于评价多肽群体中的泛素化靶标。 以这种方式，可以获得包含与多个细胞多肽的泛素化状态有关的信息的蛋白质组泛素化图。 可以获得各种不同类型的细胞和细胞状态的图。 例如，可以评估正常和患病细胞中的泛素化靶标。 优选地，地图作为数据文件存储在数据库中。 鉴定的单个泛素化多肽可用于产生诊断细胞状态的分子探针和/或可用作调节一种或多种细胞过程的试剂的靶标。

公开(公告)号：US06989129B2

公开(公告)日：2006-01-24

申请号：US10115692

申请日：2002-04-04

申请人： Lawrence J. Licklider ,  Steven P. Gygi ,  Junmin Peng

发明人： Lawrence J. Licklider ,  Steven P. Gygi ,  Junmin Peng

IPC分类号： G01N30/02 ,  G01N30/08

CPC分类号： G01N30/08 ,  G01N30/16 ,  G01N30/24 ,  G01N30/466 ,  G01N30/7266 ,  G01N2030/085 ,  G01N2030/162 ,  G01N2030/201 ,  G01N2030/202 ,  G01N2030/285 ,  Y10T436/24

摘要： A system for automatically performing liquid chromatography analysis of low volume liquid chemical samples at nanosecond flow rates using an analysis column that integrates a pre-concentration trapping column and a chromatography separation column terminating at an electrospray nozzle of an online mass spectrometer. The analysis column consists of a capillary having an inside diameter of between 75 and 125 microns packed throughout with a porous bed of micron particles. A branch outlet positioned 10 to 16 centimeters upstream from the nozzle divides the analysis column into an upstream pre-concentration trap and a downstream separation column. An autosampler delivers low volume liquid samples to the upstream inlet via a two-position valve. Feed connections couple the autosampler to upstream inlet when the valve is open to inject a liquid sample into the pre-concentration trap at a maximum loading flow rate in the range from 0.5 to 50 microliters/minute. Thereafter, when the valve closes, it terminates the further injection the sample, and a concentrated portion of the sample then passes though the chromatography separation column at a much slower flow rate between 10 and 1,000 nanoliters per minute. Throughput can be doubled by coupling two such analysis columns to a single autosampler using a ten-port, two position valve. A single column can be supplied through a six port two-position valve.