公开(公告)号：US20090299035A1

公开(公告)日：2009-12-03

申请号：US12443623

申请日：2007-10-10

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Yukiko Aruga ,  Chiori Yamane

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Yukiko Aruga ,  Chiori Yamane

IPC分类号： C07K14/00

CPC分类号： C07K14/31 ,  C07K14/315 ,  C07K17/06 ,  C07K2319/21

摘要： An object of the present invention is to provide a novel protein having the following amino acid sequence altered for specifically and efficiently binding a protein to an immobilization carrier via the carboxy terminus. The protein is used for immobilizing a portion represented by R1-R2 on an immobilization carrier, comprising the amino acid sequence represented by the general formula R1-R2-R3-R4-R5 [wherein: the sequences are oriented from the amino terminal side to the carboxy terminal side; the sequence of the R1 portion is the sequence of a subject protein to be immobilized and contains neither a lysine residue nor a cysteine residue; the sequence of the R2 portion may be absent, but when the sequence of the R2 portion is present, the sequence of the R2 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues; the sequence of the R3 portion is composed of two residues of amino acid represented by cysteine-X (where X denotes an amino acid residue other than lysine or cysteine); the sequence of the R4 portion may be absent, but when the sequence of the R4 portion is present, the sequence of the R4 portion contains neither a lysine residue nor a cysteine residue, but contains an acidic amino acid residue capable of acidifying the isoelectric point of the entire protein comprising the amino acid sequence represented by the general formula R1-R2-R3-R4-R5; and the sequence of an R5 portion is an affinity tag sequence for protein purification.

摘要翻译： 本发明的一个目的是提供一种具有以下氨基酸序列的新蛋白质，所述氨基酸序列经由羧基末端将蛋白质特异性和有效地结合到固定化载体上。 该蛋白质用于将由R1-R2表示的部分固定在固定化载体上，其包含由通式R1-R2-R3-R4-R5表示的氨基酸序列[其中：序列从氨基末端侧取向至 羧基末端侧; R1部分的序列是待固定的主题蛋白的序列，既不含有赖氨酸残基也不含有半胱氨酸残基; R2部分的序列可以不存在，但是当存在R2部分的序列时，R2部分的序列是由赖氨酸和半胱氨酸残基以外的氨基酸残基组成的间隔序列; R3部分的序列由半胱氨酸-X（其中X表示除赖氨酸或半胱氨酸以外的氨基酸残基）表示的两个氨基酸残基组成; R4部分的序列可以不存在，但当存在R4部分的序列时，R4部分的序列既不含有赖氨酸残基也不含有半胱氨酸残基，但含有能够酸化等电点的酸性氨基酸残基 包含由通式R1-R2-R3-R4-R5表示的氨基酸序列的整个蛋白质; 并且R5部分的序列是用于蛋白质纯化的亲和标签序列。

公开(公告)号：US20100130721A1

公开(公告)日：2010-05-27

申请号：US12596600

申请日：2008-04-17

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Hisashi Takahashi ,  Yukiko Aruga ,  Chiori Yamane

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Hisashi Takahashi ,  Yukiko Aruga ,  Chiori Yamane

IPC分类号： C07K17/00

CPC分类号： C07K17/00

摘要： This invention provides an immobilized protein bound to an immobilization carrier at a protein amino terminus via the sole α-amino group of the protein comprising an amino acid sequence containing neither lysine residues nor cysteine residues represented by the general formula S1-R1-R2, wherein: the sequences are oriented from the amino terminal side to the carboxy terminal side; the sequence of the S1 portion may be absent, but when the sequence of the S1 portion is present, the sequence of the S1 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues; the sequence of the R1 portion is the sequence of a subject protein to be immobilized and contains neither lysine residues nor cysteine residues; and the sequence of the R2 portion may be absent, but when the sequence of the R2 portion is present, the sequence of the R2 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues.

摘要翻译： 本发明提供了通过蛋白质的唯一α-氨基在蛋白质氨基末端与固定化载体结合的固定化蛋白，其包含不含有由通式S1-R1-R2表示的赖氨酸残基和半胱氨酸残基的氨基酸序列，其中 ：序列从氨基末端侧到羧基末端侧; S1部分的序列可以不存在，但是当存在S1部分的序列时，S1部分的序列是由赖氨酸和半胱氨酸残基以外的氨基酸残基组成的间隔序列; R1部分的序列是待固定的主题蛋白的序列，既不含有赖氨酸残基也不含有半胱氨酸残基; 并且R2部分的序列可以不存在，但是当R2部分的序列存在时，R2部分的序列是由赖氨酸和半胱氨酸残基以外的氨基酸残基组成的间隔序列。