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    Method for selectively collecting N-terminal peptide fragment of protein
    2.
    发明授权
    Method for selectively collecting N-terminal peptide fragment of protein 失效
    选择性收集蛋白质N-末端肽片段的方法

    公开(公告)号：US07041472B2

    公开(公告)日：2006-05-09

    申请号：US10739111

    申请日：2003-12-19

    申请人： Shigemi Norioka ,  Norikazu Ueyama ,  Taka-aki Okamura ,  Takashi Nakazawa ,  Minoru Yamaguchi ,  Eiji Ando

    发明人： Shigemi Norioka ,  Norikazu Ueyama ,  Taka-aki Okamura ,  Takashi Nakazawa ,  Minoru Yamaguchi ,  Eiji Ando

    IPC分类号： C12P21/06

    CPC分类号： C12P21/06

    摘要： The present invention provides a method for selectively collecting the N-terminal peptide fragments of a protein of interest whether or not the protein of interest is modified on the N-terminus. A method for selectively collecting the N-terminal peptide fragment of a protein, comprising: a protection step (1) of protecting side chain-amino groups of amino acid residues containing side chain-amino groups of a protein of interest to obtain a protected protein protected on the side chain-amino groups; a fragmentation step (2) of cleaving the protected protein into one N-terminal peptide fragment (a) containing the N-terminus of the peptide of interest and one or more of peptide fragments (b) other than the N-terminal peptide fragment (a); and a separation step (3) of separating the N-terminal peptide fragment (a) from the other peptide fragments (b) by selectively eluting the N-terminal peptide fragment (a) based on the difference in their reactivity or affinity to substrate, wherein the selective elution is achieved either by allowing the other peptide fragments (b) to bind to the substrate while allowing the N-terminal peptide fragment (a) to elute, or by allowing the N-terminal peptide fragment (a) to bind to the substrate while allowing the other peptide fragments (b) to elute and subsequently eluting the bound N-terminal peptide fragment (a).

    摘要翻译： 本发明提供了一种用于选择性收集目的蛋白质的N-末端肽片段的方法，无论目的蛋白质是否在N末端被修饰。 一种用于选择性收集蛋白质的N-末端肽片段的方法，包括：保护步骤（1）保护含有目的蛋白质的侧链 - 氨基的氨基酸残基的侧链氨基以获得受保护的蛋白质 保护侧链氨基; 将保护的蛋白质切割成包含目标肽的N末端的一个N-末端肽片段（a）和除了N-末端肽片段之外的一个或多个肽片段（b）的片段化步骤（2） 一个）; 以及通过基于它们对底物的反应性或亲和力的差异来选择性地洗脱N末端肽片段（a），从而将N-末端肽片段（a）与其它肽片段（b）分离的分离步骤（3） 其中所述选择性洗脱通过使其它肽片段（b）与底物结合同时允许N-末端肽片段（a）洗脱，或通过使N-末端肽片段（a）与 同时允许其他肽片段（b）洗脱并随后洗脱结合的N-末端肽片段（a）。

    Method for derivatizing protein or peptide to sulfonic acid derivative
    3.
    发明申请
    Method for derivatizing protein or peptide to sulfonic acid derivative 有权
    将蛋白质或肽衍生为磺酸衍生物的方法

    公开(公告)号：US20050084927A1

    公开(公告)日：2005-04-21

    申请号：US10962575

    申请日：2004-10-13

    申请人： Shigemi Norioka ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Takashi Obama ,  Eiji Ando ,  Takashi Nakazawa ,  Norikazu Ueyama ,  Taka-aki Okamura

    发明人： Shigemi Norioka ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Takashi Obama ,  Eiji Ando ,  Takashi Nakazawa ,  Norikazu Ueyama ,  Taka-aki Okamura

    IPC分类号： C07K1/107 ,  C07K1/12 ,  C07K14/78 ,  C12N9/36 ,  C12P21/06 ,  C07K14/00

    CPC分类号： C12N9/2462 ,  C07K1/1077 ,  C07K1/128 ,  C07K14/78

    摘要： The present invention provides a method for effectively introducing sulfonic acid groups into the N-terminus of a protein or a peptide; a method capable of analyzing proteins or peptides easily and effectively on mass spectrometry; an intermediate that can be used to effectively derivatize proteins or peptides as sulfonic acid derivatives. A method for derivatizing a protein or peptide to a sulfonic acid derivative, comprising steps of: modification step to react a N-terminus in a protein or peptide with a compound A which includes disulfide group, to obtain a protein or peptide modified with the compound A at the N-terminus; and cleavage step to cleave a disulfide bond of the disulfide group to convert into a sulfonic acid group, thereby converting the modified protein or peptide into a sulfonic acid derivative. A method for analyzing the amino acid sequence of a protein or peptide, wherein the sulfonic acid derivative of a protein or a peptide obtained by the above method is subjected to mass spectrometry. A protein or peptide modified with a disulfide group-containing group at the N-terminus.

    摘要翻译： 本发明提供了有效地将磺酸基引入蛋白质或肽的N末端的方法; 能够在质谱上容易且有效地分析蛋白质或肽的方法; 可用于有效地将蛋白质或肽衍生为磺酸衍生物的中间体。 一种将蛋白质或肽衍生为磺酸衍生物的方法，包括以下步骤：使蛋白质或肽中的N末端与包含二硫键的化合物A反应的修饰步骤，以获得用化合物修饰的蛋白质或肽 A在N端; 以及切割二硫键的二硫键转化为磺酸基的切割步骤，从而将修饰的蛋白质或肽转化为磺酸衍生物。 分析蛋白质或肽的氨基酸序列的方法，其中通过上述方法获得的蛋白质或肽的磺酸衍生物进行质谱分析。 在N末端用含二硫基的基团修饰的蛋白质或肽。

    Method for modifying protein or peptide C-terminal
    4.
    发明授权
    Method for modifying protein or peptide C-terminal 有权
    蛋白质或肽C端修饰方法

    公开(公告)号：US07294707B2

    公开(公告)日：2007-11-13

    申请号：US10958298

    申请日：2004-10-06

    申请人： Takashi Nakazawa ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Norikazu Ueyama ,  Taka-aki Okamura ,  Shigemi Norioka

    发明人： Takashi Nakazawa ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Norikazu Ueyama ,  Taka-aki Okamura ,  Shigemi Norioka

    IPC分类号： C07K1/00

    CPC分类号： C07K1/003

    摘要： A simple and low-cost method of selectively modifying the C-terminal of a protein or peptide is provided. A method of modifying the C-terminal of a protein or peptide comprises forming an intramolecular oxazolone ring at the C-terminal of the protein or peptide that requires C-terminal modification, and then performing a ring-opening of the oxazolone ring to produce a protein or peptide with a modified C-terminal. Preferred forms include a method in which by reacting the oxazolone ring with a compound containing a nucleophilic group to effect an oxazolone ring-opening, a protein or peptide is produced in which the C-terminal is modified with the compound containing the nucleophilic group, as well as a method in which by reacting the oxazolone ring with an active esterifying agent to effect a ring-opening, the oxazolone is converted to an active ester, which by subsequent reaction with a compound containing a nucleophilic group, produces a protein or peptide in which the C-terminal has been modified with the compound containing the nucleophilic group.

    摘要翻译： 提供了选择性修饰蛋白质或肽的C-末端的简单且低成本的方法。 修饰蛋白质或肽的C末端的方法包括在需要C末端修饰的蛋白质或肽的C-末端形成分子内的恶唑酮环，然后进行恶唑酮环的开环以产生 具有修饰C末端的蛋白质或肽。 优选的形式包括通过使恶唑酮环与含有亲核基团的化合物反应以进行恶唑酮开环的方法，制备其中C-末端用含有亲核基团的化合物修饰的蛋白质或肽作为 以及通过使恶唑酮环与活性酯化剂反应以开环的方法，将恶唑酮转化成活性酯，其随后与含有亲核基团的化合物反应，产生蛋白质或肽 其中C-末端已经与含有亲核基团的化合物一起被修饰。

    Method for modifying protein or peptide C-terminal
    5.
    发明申请
    Method for modifying protein or peptide C-terminal 有权
    蛋白质或肽C端修饰方法

    公开(公告)号：US20050085622A1

    公开(公告)日：2005-04-21

    申请号：US10958298

    申请日：2004-10-06

    申请人： Takashi Nakazawa ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Norikazu Ueyama ,  Taka-aki Okamura ,  Shigemi Norioka

    发明人： Takashi Nakazawa ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Norikazu Ueyama ,  Taka-aki Okamura ,  Shigemi Norioka

    IPC分类号： C07K1/113 ,  C07K1/00 ,  C07K1/08 ,  C07K14/00

    CPC分类号： C07K1/003

    摘要： A simple and low-cost method of selectively modifying the C-terminal of a protein or peptide is provided. A method of modifying the C-terminal of a protein or peptide comprises forming an intramolecular oxazolone ring at the C-terminal of the protein or peptide that requires C-terminal modification, and then performing a ring-opening of the oxazolone ring to produce a protein or peptide with a modified C-terminal. Preferred forms include a method in which by reacting the oxazolone ring with a compound containing a nucleophilic group to effect an oxazolone ring-opening, a protein or peptide is produced in which the C-terminal is modified with the compound containing the nucleophilic group, as well as a method in which by reacting the oxazolone ring with an active esterifying agent to effect a ring-opening, the oxazolone is converted to an active ester, which by subsequent reaction with a compound containing a nucleophilic group, produces a protein or peptide in which the C-terminal has been modified with the compound containing the nucleophilic group.

    摘要翻译： 提供了选择性修饰蛋白质或肽的C-末端的简单且低成本的方法。 修饰蛋白质或肽的C末端的方法包括在需要C末端修饰的蛋白质或肽的C-末端形成分子内的恶唑酮环，然后进行恶唑酮环的开环以产生 具有修饰C末端的蛋白质或肽。 优选的形式包括通过使恶唑酮环与含有亲核基团的化合物反应以进行恶唑酮开环的方法，制备其中C-末端用含有亲核基团的化合物修饰的蛋白质或肽作为 以及通过使恶唑酮环与活性酯化剂反应以开环的方法，将恶唑酮转化成活性酯，其随后与含有亲核基团的化合物反应，产生蛋白质或肽 其中C-末端已经与含有亲核基团的化合物一起被修饰。