Patents by Inventor Akira Tsugita
Akira Tsugita has filed for patents to protect the following inventions. This listing includes patent applications that are pending as well as patents that have already been granted by the United States Patent and Trademark Office (USPTO).
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Patent number: 8119411Abstract: The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions; In the method, a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weight are measured for theType: GrantFiled: December 20, 2010Date of Patent: February 21, 2012Assignee: NEC CorporationInventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Hiroaki Torii
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Publication number: 20110183428Abstract: The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions; In the method, a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weight are measured for theType: ApplicationFiled: December 20, 2010Publication date: July 28, 2011Applicant: NEC CORPORATIONInventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Hiroaki Torii
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Publication number: 20110053196Abstract: 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.Type: ApplicationFiled: February 8, 2007Publication date: March 3, 2011Applicant: NEC CORPORATIONInventors: Kenji Miyazaki, Yo Tabuse, Hirotaka Minagawa, Emiko Saito, Ken'ichi Kamijo, Akira Tsugita, Kozue Tsugita
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Patent number: 7879616Abstract: The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions; In the method, a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weight are measured for theType: GrantFiled: December 25, 2003Date of Patent: February 1, 2011Assignee: NEC CorporationInventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Hiroaki Torii
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Publication number: 20100167326Abstract: 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.Type: ApplicationFiled: February 8, 2007Publication date: July 1, 2010Applicant: NEC CORPORATIONInventors: Kenji Miyazaki, Ken'ichi Kamijo, Akira Tsugita, Kozue Tsugita
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Patent number: 7670841Abstract: The present invention provides, as a method of analyzing the C-terminal amino acid sequence of a peptide with use of reaction technique for successively releasing the C-terminal amino acids, in which undesirable side reactions, such as cleavage of a peptide bond at the middle of the peptide, can be prevented and chemical treatments therein can be carried out under widely applicable conditions in the course of successive release of the C-terminal amino acids from a peptide, such a method comprising steps of dehydrating the gel on which a target peptide that has been separated by gel electrophoresis is held in the bound state; immersing it in a mixture solution of an alkanoic acid anhydride added with a small amount of a perfluoroalkanoic acid in a dipolar aprotic solvent to re-swell the gel carrier, forming a 5-oxazolone structure, at a temperature chosen in the range of from 30° C. to 80° C.Type: GrantFiled: November 28, 2003Date of Patent: March 2, 2010Assignee: NEC CorporationInventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Takuji Nabetani
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Patent number: 7651859Abstract: The present invention provides, as for a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions, a following method wherein a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weightType: GrantFiled: December 4, 2003Date of Patent: January 26, 2010Assignee: NEC CorporationInventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Takuji Nabetani
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Patent number: 7588944Abstract: An analyte peptide is selectively degraded sequentially by using an alkanoic anhydride (S101). The original peptide and a series of degradation reaction products having peptide in which one or more C-terminal-sided amino acids are deleted, are subjected to a certain posttreatment (S102). The molecular weight of the reaction products is measured by mass spectrometry (S103). And, the amino acid sequence of the original peptide from C-terminal is determined, based on the molecular weight obtained by mass spectrometry (S104).Type: GrantFiled: August 24, 2004Date of Patent: September 15, 2009Assignee: NEC CorporationInventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo
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Publication number: 20090140136Abstract: The primary structure or the modification state of a protein is analyzed in detail. First, an analyte protein is subjected to PMF analysis (S101), and the gene of the protein is identified. Unidentified peaks not corresponding to the peaks of hypothetical peptide fragments are extracted, by comparing the hypothetical mass spectrum with the mass spectrum obtained by mass spectrometry of a sample protein (S102). Then, the unidentified peaks obtained are analyzed, and thus, the structure or properties of the protein, the presence and the kind of modification of amino acid residues, amino acid substitution, generation of mutants, and terminal cleavage are analyzed (S103).Type: ApplicationFiled: February 8, 2005Publication date: June 4, 2009Applicant: NEC CORPORATIONInventors: Hiroaki Torii, Kenji Miyazaki, Kenichi Kamijo, Akira Tsugita
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Publication number: 20080213911Abstract: An analyte peptide is selectively degraded sequentially by using an alkanoic anhydride (S101). The original peptide and a series of degradation reaction products having peptide in which one or more C-terminal-sided amino acids are deleted, are subjected to a certain posttreatment (S102). The molecular weight of the reaction products is measured by mass spectrometry (S103). And, the amino acid sequence of the original peptide from C-terminal is determined, based on the molecular weight obtained by mass spectrometry (S104).Type: ApplicationFiled: August 24, 2004Publication date: September 4, 2008Inventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo
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Patent number: 7384790Abstract: The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by applying reaction technique for successively releasing the C-terminal amino acids therefrom, which method can suppress, when releasing the C-terminal amino acids of the peptide in sequence, such as an undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide, and allows to carry out the chemical treatment thereof under widely applicable conditions. In the method according to the present invention, an alkanoic acid anhydride and a perfluoroalkanoic acid both of vapor phase, which are supplied from a mixture containing an alkanoic acid anhydride with a small amount of a perfluoroalkanoic acid added thereto, are allowed to act on a dry sample of the peptide to be examined in a dry atmosphere at a temperature chosen in a range of 15 to 60° C.Type: GrantFiled: March 24, 2003Date of Patent: June 10, 2008Assignee: NEC CorporationInventors: Kenji Miyazaki, Akira Tsugita, Naoyuki Takahashi, Takuji Nabetani, Toshimasa Yamazaki, Kenichi Kamijo
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Publication number: 20070148720Abstract: The present invention provides an approach for identifying with high accuracy, a known protein or a variant of the known protein derived from the same genomic gene as in a target protein to be analyzed, based on a mass spectrometric result of a plurality of peptide fragments obtained from site-specific enzymatic digestion of the target protein to be analyzed by referring to nucleotide sequences of genes encoding known proteins on a database and to deduced full-length amino acid sequences thereof.Type: ApplicationFiled: December 17, 2004Publication date: June 28, 2007Inventors: Hiroaki Torii, Kenji Miyazaki, Kenichi Kamijo, Akira Tsugita
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Patent number: 7112446Abstract: In a method for efficiently analyzing a posttranslational modification of a protein using no enzyme, a protein or peptide to be analyzed is reacted with an acid (a thioester or hydrazine) under certain conditions. This makes it possible to detect variously modified states of a protein or peptide, whereby the identification of each specific modifying group and the position of each modified amino acid can be efficiently analyzed using a chemical method and a mass spectrometric apparatus.Type: GrantFiled: March 14, 2002Date of Patent: September 26, 2006Assignees: Tokyo Rikakikai Co., Ltd., NEC CorporationInventors: Hirotaka Minagawa, Ichiro Ishida, Akira Tsugita, Takuji Nabetani
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Publication number: 20060177887Abstract: Provided a method of acquiring information on internal sequence which is applicable to a trace amount of protein and has a high reliability. A protein to be analyzed is limitedly cleaved at a specified position of amino acid to prepare a mixture of a plurality of peptide fragments; one or more peptide fragments are isolated and purified from the peptide fragment mixture; C-terminal amino acids of the isolated and purified peptide fragment are successively released by chemical reaction to prepare a mixture containing a series of resulting products; mass spectrometry is applied to both the reaction products of the successive truncation and unreacted peptide fragment not subjected to the successive truncation reaction; then results of the mass spectrometry is analyzed to acquire chemical structure information of the target protein.Type: ApplicationFiled: January 3, 2006Publication date: August 10, 2006Inventors: Kenji Miyazaki, Hiroaki Torii, Kenichi Kamijo, Akira Tsugita
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Publication number: 20060134720Abstract: The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions; In the method, a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weight are measured for theType: ApplicationFiled: December 25, 2003Publication date: June 22, 2006Inventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Hiroaki Torii
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Publication number: 20060057731Abstract: The present invention provides, as for a method for analyzing the C-terminal amino acid sequence of a peptide by using a reaction for successively releasing the C-terminal amino acids of the peptide, which method can suppress, when successively releasing the C-terminal amino acids of a peptide of long amino acid length, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide and can carry out the chemical treatment thereof under widely applicable conditions, a following method wherein a dry sample of a peptide with long amino acid length is beforehand subjected to an N-acylation treatment; by using a reaction reagent where an alkanoic acid anhydride is combined with a small amount of a perfluoroalkanoic acid, successive release of C-terminal amino acids is conducted under mild conditions; a hydrolysis treatment is applied; then, selective fragmentization at site of arginine residue is performed by digestion by trypsin; thereafter, decreases in molecular weightType: ApplicationFiled: December 4, 2003Publication date: March 16, 2006Applicants: NEC Corporation, Tokyo Rikakikai Co., Ltd.Inventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Takuji Nabetani
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Publication number: 20060030052Abstract: The present invention provides, as a method of analyzing the C-terminal amino acid sequence of a peptide with use of reaction technique for successively releasing the C-terminal amino acids, in which undesirable side reactions, such as cleavage of a peptide bond at the middle of the peptide, can be prevented and chemical treatments therein can be carried out under widely applicable conditions in the course of successive release of the C-terminal amino acids from a peptide, such a method comprising steps of dehydrating the gel on which a target peptide that has been separated by gel electrophoresis is held in the bound state; immersing it in a mixture solution of an alkanoic acid anhydride added with a small amount of a perfluoroalkanoic acid in a dipolar aprotic solvent to re-swell the gel carrier, forming a 5-oxazolone structure, at a temperature chosen in the range of from 30° C. to 80° C.Type: ApplicationFiled: November 28, 2003Publication date: February 9, 2006Inventors: Kenji Miyazaki, Akira Tsugita, Kenichi Kamijo, Takuji Nabetani
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Publication number: 20050074832Abstract: The present invention provides a method for analyzing the C-terminal amino acid sequence of a peptide by applying reaction technique for successively releasing the C-terminal amino acids therefrom, which method can suppress, when releasing the C-terminal amino acids of the peptide in sequence, such a undesirable side reaction as cleavage of peptide bond in the intermediate position of the peptide, and allows to carry out the chemical treatment thereof under widely applicable conditions. In the method according to the present invention, an alkanoic acid anhydride and a perfluoroalkanoic acid both of vapor phase, which are supplied from a mixture containing an alkanoic acid anhydride with a small amount of a perfluoroalkanoic acid added thereto, are allowed to act on a dry sample of the peptide to be examined in a dry atmosphere at a temperature chosen in a range of 15 to 60° C.Type: ApplicationFiled: March 24, 2003Publication date: April 7, 2005Inventors: Kenji Miyazaki, Akira Tsugita, Naoyuki Takahashi, Takuji Nabetani, Toshimasa Yamazaki, Kenichi Kamijo
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Publication number: 20040014229Abstract: In a method for efficiently analyzing a posttranslational modification of a protein using no enzyme, a protein or peptide to be analyzed is reacted with an acid (a thioester or hydrazine) under certain conditions. This makes it possible to detect variously modified states of a protein or peptide, whereby the identification of each specific modifying group and the position of each modified amino acid can be efficiently analyzed using a chemical method and a mass spectrometric apparatus.Type: ApplicationFiled: March 14, 2002Publication date: January 22, 2004Applicants: NEC CORPORATION, TOKYO RIKAKIKAI CO., LTD.Inventors: Hirotaka Minagawa, Ichiro Ishida, Akira Tsugita, Takuji Nabetani
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Patent number: 6046053Abstract: A process is disclosed for sequencing proteins or peptides from the C-terminal end. The process comprises the steps of reacting the peptide or protein with an alkyl acid anhydride to convert the carboxy-terminal thereof into oxazolone, liberating the C-terminal amino acid by reaction with acid and alcohol or with ester, and identifying the liberated amino acid or amino acid derivative.Type: GrantFiled: May 23, 1997Date of Patent: April 4, 2000Assignee: Seiko Instruments Inc.Inventors: Akira Tsugita, Keiji Takamoto, Tatsuaki Ataka, Toshihiko Sakuhara, Toyoaki Uchida