公开(公告)号：US07579165B2

公开(公告)日：2009-08-25

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： C07K1/13 ,  C07K1/14 ,  C07K14/00 ,  C12P21/06 ,  C12Q1/00 ,  C12Q1/34 ,  C12Q1/37 ,  G01N33/53 ,  G01N33/38 ,  G01N33/483

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. The method comprises a modification step to react a N-terminus in a protein or peptide with a compound A which includes disulfide group and a cleavage step to cleave a disulfide bond of the disulfide group to convert into a sulfonic acid group. The present invention also provides a method of analyzing proteins or peptides easily and effectively on mass spectrometry and an intermediate that can be used to effectively derivatize proteins or peptides as sulfonic acid derivatives.

摘要翻译： 本发明提供了将磺酸基团有效地引入蛋白质或肽的N-末端的方法。 该方法包括使蛋白质或肽中的N末端与包含二硫键的化合物A和切割二硫键的二硫键以切割成磺酸基团的切割步骤进行反应的修饰步骤。 本发明还提供了一种在质谱分析中容易且有效地分析蛋白质或肽的方法，以及可用于有效地将蛋白质或肽衍生为磺酸衍生物的中间体。

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。

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。

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。

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）。

公开(公告)号：US20090325227A1

公开(公告)日：2009-12-31

申请号：US12090747

申请日：2006-10-23

申请人： Takashi Nakazawa ,  Mutsumi Oka ,  Kimiko Nishida ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Norikazu Ueyama ,  Takaaki Okamura ,  Susumu Tsunasawa

发明人： Takashi Nakazawa ,  Mutsumi Oka ,  Kimiko Nishida ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Norikazu Ueyama ,  Takaaki Okamura ,  Susumu Tsunasawa

IPC分类号： C12P21/00 ,  C07K1/107 ,  G01N33/68

CPC分类号： C07K17/00 ,  C07K1/1072 ,  C07K1/113

摘要： The present invention provides a method for inexpensively, easily and efficiently modifying the C-terminus of a protein or peptide; a method for easily and reliably isolating a C-terminal peptide fragment of a protein or peptide; and a method for rapidly, accurately and reliably determining an amino acid sequence of a protein or peptide by using a mass spectroscope. A method comprising the step of adding a formylation reagent and a catalyst to a protein or peptide to convert a carboxyl group into an aldehyde group. A method comprising the step of reacting a nucleophilic reagent with the aldehyde group to modify the C-terminus of the protein or peptide. A method comprising the step of reacting a support having a nucleophilic group to immobilize the protein or peptide. A method comprising the step of fragmenting the immobilized protein or peptide, washing the support, and isolating the C-terminal peptide fragment from the support. A method comprising the step of subjecting the isolated C-terminal peptide fragment to mass spectrometry and determining an amino acid sequence.

摘要翻译： 本发明提供了一种廉价，容易且有效地修饰蛋白质或肽的C末端的方法; 容易且可靠地分离蛋白质或肽的C末端肽片段的方法; 以及通过使用质谱仪快速，准确，可靠地测定蛋白质或肽的氨基酸序列的方法。 一种方法，其包括将甲酰化试剂和催化剂加入到蛋白质或肽中以将羧基转化为醛基的步骤。 一种包括使亲核试剂与醛基反应以修饰蛋白质或肽的C末端的步骤的方法。 一种包括使具有亲核基团的载体反应以固定蛋白质或肽的方法。 一种方法，其包括将固定的蛋白质或肽片段化，洗涤载体和从载体中分离C末端肽片段的步骤。 一种方法，包括使分离的C-末端肽片段进行质谱分析和测定氨基酸序列的步骤。

公开(公告)号：US20090142851A1

公开(公告)日：2009-06-04

申请号：US12325115

申请日：2008-11-28

申请人： Keisuke Shima ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Osamu Nishimura ,  Susumu Tsunasawa ,  Kazuhiro Sonomura

发明人： Keisuke Shima ,  Minoru Yamaguchi ,  Hiroki Kuyama ,  Eiji Ando ,  Osamu Nishimura ,  Susumu Tsunasawa ,  Kazuhiro Sonomura

IPC分类号： G01N33/68 ,  C07K1/14 ,  C12P21/06

CPC分类号： C07K1/1072 ,  C07K1/128 ,  G01N33/6821

摘要： The present invention provides a method for specifically recovering a C-terminal peptide fragment, and a method for easily determining the sequence of a C-terminal peptide fragment, which is difficult to be determined by a conventional method, with the use of a mass spectrometer, in particular a method capable of de novo sequencing of a C-terminal peptide fragment. A method for selectively recovering a C-terminal peptide of a protein, comprising the steps of: in a cleavage product of a protein containing a C-terminal peptide fragment (A) having an α-amino group but not having an ε-amino group and the other peptide fragments (B) having an α-amino group and an ε-amino group, selectively modifying the α-amino groups to obtain a C-terminal peptide fragment modified (A′) and the other peptide fragments modified (B′); and separating the C-terminal peptide fragment modified (A′) from the modified cleavage product by allowing a carrier to hold the other peptide fragments modified (B′) via the ε-amino group. A method for determining the amino acid sequence of a C-terminal peptide of a protein, comprising the steps of: selectively recovering a C-terminal peptide of a protein by the above method; and determining the amino acid sequence by subjecting a recovered C-terminal peptide fragment to mass spectrometry measurement.

摘要翻译： 本发明提供了特异性回收C末端肽片段的方法，以及通过使用质谱仪容易地测定难以用常规方法测定的C末端肽片段的序列的方法 ，特别是能够重新测序C-末端肽片段的方法。 一种选择性回收蛋白质C末端肽的方法，包括以下步骤：在含有具有α-氨基但不具有ε-氨基的C-末端肽片段（A）的蛋白质的切割产物中 和具有α-氨基和ε-氨基的其它肽片段（B），选择性地修饰α-氨基以获得修饰的C末端肽片段（A'）和修饰的其它肽片段（B' ）; 以及通过允许载体通过ε-氨基保持修饰的其它肽片段（B'），从修饰的切割产物中分离修饰的C末端肽片段（A'）。 一种确定蛋白质C-末端肽的氨基酸序列的方法，包括以下步骤：通过上述方法选择性地回收蛋白质的C-末端肽; 并通过对回收的C-末端肽片段进行质谱测量来测定氨基酸序列。

公开(公告)号：US20050221413A1

公开(公告)日：2005-10-06

申请号：US10518358

申请日：2003-04-03

申请人： Hiroki Kuyama ,  Eiji Ando ,  Osamu Nishimura

发明人： Hiroki Kuyama ,  Eiji Ando ,  Osamu Nishimura

IPC分类号： C07C313/08 ,  G01N33/68 ,  C12Q1/37 ,  C07K14/00 ,  G01N33/00

CPC分类号： G01N33/6848 ,  C07B2200/05 ,  C07C313/08 ,  G01N2458/15

摘要： A sulfenyl compound represented by the general formula: R—S—X (I)(wherein R represents an organic group having at least one constituent element labeled with an isotope, and X represents a leaving group); a labeling reagent comprising it; and a method of analyzing peptide using the labeling reagent. Preferably the organic group R comprises C, H, and N, and optionally O and/or P as the constituent element, and the isotope is a stable isotope selected from the group consisting of 2H, 13C, 15N, 17O, and 18O.

摘要翻译： 由通式R-S-X（I）（其中R表示具有至少一个由同位素标记的构成元素，X表示离去基团的有机基团）表示的亚磺酰基化合物; 包含它的标记试剂; 以及使用标记试剂分析肽的方法。 优选地，有机基团R包含C，H和N，以及任选的O和/或P作为构成元素，并且同位素是选自下组的稳定同位素：SUP， > 13℃，15℃，17℃和<！ - SIPO - > O。

公开(公告)号：US09018332B2

公开(公告)日：2015-04-28

申请号：US12863165

申请日：2009-01-14

申请人： Tadashi Okawa ,  Eiji Ando

发明人： Tadashi Okawa ,  Eiji Ando

IPC分类号： C08G77/20 ,  C08G77/38 ,  C08G77/12 ,  C08G77/46

CPC分类号： C08G77/38 ,  C08G77/12 ,  C08G77/46

摘要： A methacryloxy group- or acryloxy group-containing polyorganosiloxane in which a methacryloxy group or acryloxy group is bonded to a silicon atom in the polyorganosiloxane across a long-chain alkylene group or poly(alkyleneoxy)-long chain-alkylene group having 11 to 20 carbon atoms. A method of producing this methacryloxy group- or acryloxy group-containing polyorganosiloxane, in which a silicon-bonded hydrogen atom-containing polyorganosiloxane is addition reacted in the presence of a hydrosilylation reaction catalyst with a 1-alkenyl methacrylate or a 1-alkenyl acrylate or a 1-alkenyloxypolyalkylene glycol methacrylate or a 1-alkenyloxypolyalkylene glycol acrylate.

摘要翻译： 含有甲基丙烯酰氧基或丙烯酰氧基的聚有机硅氧烷，其中甲基丙烯酰氧基或丙烯酰氧基通过长链亚烷基或聚（亚烷氧基） - 长链亚烷基键合到聚有机硅氧烷中的硅原子上，其具有11至20个碳 原子 在含硅键合的含氢原子的聚有机硅氧烷的加成反应催化剂的存在下，在氢化硅烷化反应催化剂与1-链烯基甲基丙烯酸酯或1-烯基丙烯酸酯的情况下生产该含甲基丙烯酰氧基或含丙烯酰氧基的聚有机硅氧烷的方法，或 1-链烯氧基聚亚烷基二醇甲基丙烯酸酯或1-链烯氧基聚亚烷基二醇丙烯酸酯。

公开(公告)号：US20120186916A1

公开(公告)日：2012-07-26

申请号：US13440002

申请日：2012-04-05

申请人： Hiroshi Kigawa ,  Naoyuki Maruyama ,  Eiji Ando

发明人： Hiroshi Kigawa ,  Naoyuki Maruyama ,  Eiji Ando

IPC分类号： B66B5/02

CPC分类号： B66B5/00

摘要： In a safety system for an elevator, slip detection means detects a slip between a drive sheave and a main rope. A safety gear is mounted to a car, the safety gear being electrically operated by an actuator to cause the car to make an emergency stop regardless of whether a running direction of the car is upward or downward. A safety gear controller cuts power supply to a hoisting machine motor and causes the safety gear to make a braking operation upon detection of the slip between the drive sheave and the main rope by the slip detection means.

摘要翻译： 在电梯的安全系统中，滑动检测装置检测驱动绳轮和主绳索之间的滑动。 安全装置安装在汽车上，安全钳由致动器电动操作，使轿厢进行紧急停止，而不管汽车的行驶方向是向上还是向下。 安全钳子控制器切断曳引机马达的电源，并通过滑动检测装置检测到驱动绳轮和主绳索之间的滑动，使安全装置进行制动操作。