公开(公告)号：US20100167326A1

公开(公告)日：2010-07-01

申请号：US12278878

申请日：2007-02-08

申请人： Kenji Miyazaki ,  Ken'ichi Kamijo ,  Akira Tsugita ,  Kozue Tsugita

发明人： Kenji Miyazaki ,  Ken'ichi Kamijo ,  Akira Tsugita ,  Kozue Tsugita

IPC分类号： C12Q1/37 ,  C07K1/107 ,  G01N33/00

CPC分类号： C07K1/107 ,  C07K1/128 ,  G01N33/6821 ,  G01N33/6848

摘要： The present invention is to provide a method for cleavage of peptidic bond at C terminal of peptide and a method for determination of C terminal amino acid sequence of peptide without the decrease of the sensitivity and with preventing the subsidiary reaction. The method for cleavage of peptidic bond at C terminal of peptide according to the present invention is characterized as follows: A method for cleavage of peptidic bond at C terminal of peptide comprising the steps of: reacting a peptide with a solution of perhalogenated carboxylic acid containing alcohol to esterify glutamic acid residue of said peptide; and reacting said peptide with alkanoic acid anhydride to obtain C terminal-deleted peptide in which the amino acid residue of said peptide at C terminal is sequentially deleted. The method for determination of C terminal amino acid sequence of peptide according to the present invention is characterized as follows: A method for determination of C terminal amino acid sequence of peptide comprising the steps of: reacting a peptide with a solution of perhalogenated carboxylic acid containing alcohol to esterify glutamic acid residue of said peptide; reacting said peptide with alkanoic acid anhydride to obtain C terminal-deleted peptide in which the amino acid residue of said peptide at C terminal is sequentially deleted; measuring molecular weight of said C terminal-deleted peptide; and grasping said peptide based on said molecular weight.

摘要翻译： 本发明提供肽的C末端肽键的切割方法以及肽的C末端氨基酸序列的测定方法，而不会降低灵敏度并防止辅助反应。 根据本发明的肽在C末端切割肽键的方法的特征如下：肽的C末端切割肽键的方法，其包括以下步骤：使肽与含有全卤代羧酸的溶液反应 醇以酯化所述肽的谷氨酸残基; 并使所述肽与链烷酸酐反应，得到C末端缺失的肽，其中C末端的所述肽的氨基酸残基被顺序删除。 根据本发明的肽的C末端氨基酸序列的测定方法的特征如下：肽的C末端氨基酸序列的测定方法，包括以下步骤：使肽与含有卤素的全卤代羧酸的溶液反应 醇以酯化所述肽的谷氨酸残基; 使所述肽与链烷酸酐反应，得到C末端缺失的肽，其中所述肽在C末端的氨基酸残基被顺序删除; 测量所述C末端缺失肽的分子量; 并基于所述分子量掌握所述肽。

公开(公告)号：US20110053196A1

公开(公告)日：2011-03-03

申请号：US12278711

申请日：2007-02-08

申请人： Kenji Miyazaki ,  Yo Tabuse ,  Hirotaka Minagawa ,  Emiko Saito ,  Ken'ichi Kamijo ,  Akira Tsugita ,  Kozue Tsugita

发明人： Kenji Miyazaki ,  Yo Tabuse ,  Hirotaka Minagawa ,  Emiko Saito ,  Ken'ichi Kamijo ,  Akira Tsugita ,  Kozue Tsugita

IPC分类号： C12Q1/37 ,  C07K1/113

CPC分类号： G01N33/6803 ,  C07K1/107

摘要： The present invention is to provide a method for easily and specifically modifying specific amino acid residue(s) constituting a peptide and to provide a methodology of improving the accuracy of identification of the peptide using a new information of the peptide obtained from the number of modified amino acid residue by said specific modification method as mentioned. The method for modifying a peptide according to the present invention is characterized:A method for modifying a peptide, wherein the peptide as supported in a substrate and an aqueous solution of perhalogenated carboxylic acid containing an alcohol is reacted to selectively esterify a glutamic acid residue of said peptide.The method for identifying a peptide according to the present invention is characterized:A method for identifying a peptide comprising the steps of: reacting the peptide as supported in a substrate and an aqueous solution of perhalogenated carboxylic acid containing an alcohol to selectively esterify glutamic acid residue of said peptide; immersing said substrate in a protease solution to obtain a peptide fragment originated from said peptide; measuring a molecular weight of said peptide fragment; and determining said peptide based on said molecular weight.

摘要翻译： 本发明提供一种容易且特异性地修饰构成肽的特定氨基酸残基的方法，并且提供使用从修饰的数量获得的肽的新信息来提高肽的鉴定准确度的方法 通过所述具体的改性方法制备氨基酸残基。 本发明的肽的修饰方法的特征在于：对肽进行修饰的方法，其中负载在底物中的肽和含有醇的全卤代羧酸的水溶液进行反应以选择性地酯化谷氨酸残基 所述肽。 本发明鉴定肽的方法的特征在于：一种鉴定肽的方法，包括以下步骤：将载体载于底物中的肽与含有醇的全卤代羧酸水溶液反应以选择性酯化谷氨酸残基 的所述肽; 将所述底物浸入蛋白酶溶液中以获得源自所述肽的肽片段; 测量所述肽片段的分子量; 并基于所述分子量测定所述肽。