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    • 4. 发明申请
    • Method for measuring protein NMR
    • 蛋白质NMR测定方法
    • US20060057660A1
    • 2006-03-16
    • US11199192
    • 2005-08-09
    • Yaeta EndouToshiyuki KohnoHayato Morita
    • Yaeta EndouToshiyuki KohnoHayato Morita
    • C12Q1/37C12P21/06
    • G01N24/08G01R33/465
    • An object of the present invention is to provide a means for highly efficiently or quickly evaluating or determining the three-dimensional structure of a protein by conducting multidimensional multinuclear NMR measurement without performing a step of separating and purifying the synthesized labeled protein from other labeled proteins. The present invention provides a method for analyzing proteins, which comprises: synthesizing a protein of interest under conditions in which 90% or more of the synthesized labeled proteins constitute the protein of interest, when proteins are synthesized using a nucleic acid encoding the protein of interest as a template and also using substrate amino acids, a part or all of which have been labeled in such a way that NMR measurement is possible; and performing the NMR measurement using the above-described labeled proteins.
    • 本发明的目的是提供通过进行多维多核NMR测量来高效或快速地评估或确定蛋白质的三维结构的方法,而不进行从其它标记的蛋白质分离和纯化合成的标记蛋白质的步骤。 本发明提供了一种分析蛋白质的方法,其包括:当使用编码感兴趣的蛋白质的核酸合成蛋白质时,在其中90%或更多的合成的标记蛋白质构成感兴趣的蛋白质的条件下合成感兴趣的蛋白质 作为模板并且还使用底物氨基酸,其部分或全部已经以可测量NMR测量的方式被标记; 并使用上述标记的蛋白进行NMR测定。
    • 5. 发明授权
    • Rotary piston internal combustion engine
    • 旋转活塞内燃机
    • US4003346A
    • 1977-01-18
    • US514906
    • 1974-10-15
    • Toshiyuki Kohno
    • Toshiyuki Kohno
    • F01C1/22F02B53/00F02B55/16F02B53/04
    • F01C1/22F02B55/16F02B2053/005F02B53/00Y02T10/17
    • A rotary engine having a housing defining an internal rotor chamber between a trochoidally shaped peripheral wall and a pair of side walls is provided with a peripheral port through which a rich air-fuel mixture is supplied, and a side port, through which air is supplied, with the side port being arranged forwardly of the peripheral port taken in the direction of rotor rotation. The peripheral port opens into the combustion chamber of the engine through the peripheral wall and the side port opens through one of the side walls. The degree of opening of the side port changes in accordance with rotor rotation and when the rotor is within a certain angular range following the top dead center position during the intake stroke of its rotational cycle, both ports are open into the combustion chamber defined between the housing and the rotor.
    • 一种具有壳体,该壳体限定了位于齿状周缘壁与一对侧壁之间的内部转子室,该外壳设置有供给富空燃混合物的外围端口和供给空气的侧端口 侧端口被设置在沿转子旋转方向截取的外围端口的前方。 外围端口通过周壁进入发动机的燃烧室,侧端口通过一个侧壁打开。 侧端口的开度根据转子旋转而变化,并且当转子在其旋转周期的进气冲程期间在上止点位置之后处于一定角度范围内时,两个口都打开到在 外壳和转子。
    • 6. 发明申请
    • Nmr Signal Assignment Method
    • Nmr信号分配方法
    • US20080293150A1
    • 2008-11-27
    • US10588152
    • 2005-02-02
    • Toshiyuki Kohno
    • Toshiyuki Kohno
    • G01N33/68
    • G01N24/08G01R33/465
    • It is an object of the present invention to provide a method of efficiently and rapidly determining the assignment of signals obtained by the 1H-15N HSQC measurement or the like, and a method of efficiently and rapidly identifying the three-dimensional structure of a target protein using the above method or a method of identifying the binding site of a protein of interest to a ligand, in place of the conventional signal assignment method, which requires a protein with a concentration several to 10 times higher than the lowest concentration of a protein capable of being observed by a high sensitive 1H-15N HSQC measurement method. In the present invention, 15N/13C double-labeled amino acids, 15N-labeled amino acids, and non-labeled amino acids are systematically combined and used as substrates with respect to every amino acids constituting a protein of interest, so as to synthesize multiple proteins (20 types or 39 types of proteins at the maximum), and the NMR measurement is performed on such multiple proteins by a measurement method capable of identifying the correlation signals of two amino acid residues adjacent to each other, and then the obtained signals are compared.
    • 本发明的目的是提供一种有效和快速地确定通过1H-15N HSQC测量等获得的信号的分配的方法,以及有效和快速地鉴定靶蛋白的三维结构的方法 使用上述方法或鉴定目的蛋白质与配体的结合位点的方法代替传统的信号分配方法,其需要的浓度高于最低浓度的蛋白质的几十倍的蛋白质 通过高灵敏度的1H-15N HSQC测量方法观察到。 在本发明中,将15N / 13C双标氨基酸,15N标记氨基酸和非标记氨基酸系统地组合并用作相对于构成目的蛋白质的每个氨基酸的底物,以便合成多重 蛋白质(最多20种或39种蛋白质),并且通过能够鉴定彼此相邻的两个氨基酸残基的相关信号的测量方法对这样的多种蛋白进行NMR测定,然后获得的信号为 比较。
    • 8. 发明申请
    • Method for synthesizing amino-acid-selectively labeled protein
    • 合成氨基酸选择性标记蛋白的方法
    • US20060216791A1
    • 2006-09-28
    • US11340800
    • 2006-01-27
    • Yaeta EndouToshiyuki Kohno
    • Yaeta EndouToshiyuki Kohno
    • C12P21/04
    • C12P21/02G01N33/6848
    • An object of the present invention is to provide a method for synthesizing, in a cell-free protein synthetic system using a wheat germ extract, a protein of interest that is labeled in an amino-acid-selective manner. The present invention provides a method for carrying out a translation reaction of a protein of interest under a condition whereby a metabolic reaction of alanine to aspartic acid and glutamic acid, a metabolic reaction of aspartic acid to glutamic acid or a metabolic reaction of glutamic acid to aspartic acid and/or glutamine is inhibited, while the protein synthesis is not inhibited, when alanine, aspartic acid or glutamic acid of the protein of interest is labeled with a stable isotope in a cell-free protein synthetic system containing a wheat germ extract.
    • 本发明的目的是提供一种在使用小麦胚提取物的无细胞蛋白质合成系统中合成以氨基酸选择性方式标记的感兴趣的蛋白质的方法。 本发明提供了一种在丙氨酸与天冬氨酸和谷氨酸的代谢反应,天冬氨酸对谷氨酸的代谢反应或谷氨酸的代谢反应到谷氨酸的代谢反应的条件下进行感兴趣的蛋白质的翻译反应的方法 当蛋白质的丙氨酸,天冬氨酸或谷氨酸在含有小麦胚芽提取物的无细胞蛋白质合成系统中用稳定同位素标记时,天冬氨酸和/或谷氨酰胺被抑制,而蛋白质合成不被抑制。