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    Novel n-acetylgalactosamine transferases and nucleic acids encoding the same
    32.
    发明申请
    Novel n-acetylgalactosamine transferases and nucleic acids encoding the same 失效
    新型的N-乙酰半乳糖胺转移酶和编码相同的核酸

    公开(公告)号：US20060234232A1

    公开(公告)日：2006-10-19

    申请号：US10524505

    申请日：2003-08-13

    申请人： Hisashi Narimatsu ,  Masanori Gotoh ,  Takashi Sato

    发明人： Hisashi Narimatsu ,  Masanori Gotoh ,  Takashi Sato

    IPC分类号： C12Q1/68 ,  C07H21/04 ,  C12P21/06 ,  C12N9/10

    CPC分类号： C12N9/1051 ,  G01N33/57484

    摘要： An enzyme which transfers N-acetylgalactosamine to N-acetylglucosamine via a β1-4 linkage was isolated and the structure of its gene was explained. This led to the production of said enzyme or the like by genetic engineering techniques, the production of oligosaccharides using said enzyme, and the diagnosis of diseases on the basis of said gene or the like. The present invention uses a protein having the amino acid sequence shown in SEQ ID NO: 1, 3, 26 or 27 in the Sequence Listing or a variant of said amino acid sequence wherein one or more acids are substituted or deleted, or one or more acids are inserted or added and having the activity of transferring N-acetylgalactosamine (GalNAc) to N-acetylglucosamine serving as a substrate via a β1-4 linkage and nucleic acids encoding said protein.

    摘要翻译： 分离通过β1-4键将N-乙酰半乳糖胺转移到N-乙酰葡糖胺的酶，并解释其基因的结构。 这导致通过基因工程技术生产所述酶等，使用所述酶生产寡糖，以及基于所述基因等诊断疾病。 本发明使用具有序列表中SEQ ID NO：1,3,26或27所示氨基酸序列的蛋白质或其中一个或多个酸被取代或缺失的所述氨基酸序列的变体，或一个或多个 酸被插入或添加并且具有通过β1-4键和编码所述蛋白质的核酸将N-乙酰半乳糖胺（GalNAc）转移到用作底物的N-乙酰葡糖胺的活性。

    Novel n-acetylglucosamine transferase, nucleic acid encoding the same and use thereof in diagnosing cancer and/or tumor
    33.
    发明申请
    Novel n-acetylglucosamine transferase, nucleic acid encoding the same and use thereof in diagnosing cancer and/or tumor 有权
    新的正乙酰葡糖胺转移酶，编码相同的核酸和其在诊断癌症和/或肿瘤中的用途

    公开(公告)号：US20050186570A1

    公开(公告)日：2005-08-25

    申请号：US10507421

    申请日：2003-03-14

    申请人： Hisashi Narimatsu ,  Niro Inaba ,  Akira Togayachi

    发明人： Hisashi Narimatsu ,  Niro Inaba ,  Akira Togayachi

    IPC分类号： C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N9/10 ,  C12N15/12 ,  C12Q1/68 ,  C07H21/04

    CPC分类号： C12N9/1051 ,  C12Q1/6886 ,  C12Q2600/158

    摘要： An enzyme having an activity of transferring N-acetylglucosamine to the non-reducing end of a Gal β1-4Glc or Gal β1-4GlcNAc-group via a β-1,3 bond; a nucleic acid encoding the same; and a method of diagnosing cancer and/or tumor, in particular, digestive cancer and/or tumor using the expression dose of a gene of the above enzyme as an indication. A gene of a novel enzyme having an activity of transferring N-acetylglucosamine to the non-reducing end of a Gal β1-4Glc or Gal β1-4GlcNAc-group via a β-1,3 bond is cloned from human stomach cells and its base sequence is determined. Then this enzyme is expressed. Since this enzyme is scarcely or never produced in cancer and/or tumor, in particular, digestive cancer and/or tumor cells, cancer and/or tumor can be diagnosed with the use of the expression of the enzyme gene as an indication.

    摘要翻译： 具有通过β-1,3键将N-乙酰葡糖胺转移至Galβ1-4Glc或Galβ1-4GlcNAc-基团的非还原末端的活性的酶; 编码该核酸的核酸; 以及使用上述酶的基因的表达剂量作为指标来诊断癌症和/或肿瘤，特别是消化癌和/或肿瘤的方法。 从人胃细胞及其碱基克隆具有通过β-1,3键将N-乙酰葡糖胺转移至Galβ1-4Glc或Galβ1-4GlcNAc-基团的非还原端的活性的新型酶的基因 确定序列。 然后表达这种酶。 由于这种酶在癌症和/或肿瘤中几乎没有或从未产生，特别是消化癌和/或肿瘤细胞，所以可以用酶基因的表达作为指示诊断癌症和/或肿瘤。

    Method for producing sialic-acid-containing sugar chain
    35.
    发明授权
    Method for producing sialic-acid-containing sugar chain 有权
    含唾液酸糖链的方法

    公开(公告)号：US09376506B2

    公开(公告)日：2016-06-28

    申请号：US14002645

    申请日：2012-02-27

    申请人： Yasunori Chiba ,  Yoshie Takahashi ,  Hisashi Narimatsu ,  Kazuhiro Fukae

    发明人： Yasunori Chiba ,  Yoshie Takahashi ,  Hisashi Narimatsu ,  Kazuhiro Fukae

    IPC分类号： C12P19/18 ,  C12N15/09 ,  C12N15/52 ,  C12P19/04 ,  C08B37/00 ,  C12P21/00

    CPC分类号： C08B37/0063 ,  C08B37/006 ,  C12N9/1081 ,  C12N15/09 ,  C12N15/52 ,  C12P19/04 ,  C12P19/18 ,  C12P19/26 ,  C12P21/005 ,  C12Y204/99 ,  C12Y204/99001 ,  C12Y301/03

    摘要： [Problem to be Solved]The importance of sugar chains having α2,3- or α2,6-linked sialic acid at their non-reducing ends is known. Industrial production has been demanded for these sugar chain compounds. Particularly, the production of glycoprotein drugs or the like inevitably requires producing in quantity sugar chains having homogeneous structures by controlling the linking pattern (α2,6-linkage or α2,3-linkage) of sialic acid. Particularly, a triantennary or tetraantennary N-type complex sugar chain having sialic acid at each of all non-reducing ends is generally considered difficult to chemically synthesize. There has been no report disclosing that such a sugar chain was chemically synthesized. Furthermore, these sugar chains are also difficult to efficiently prepare enzymatically.[Solution]The present inventors have newly found the activity of sialyltransferase of degrading sialic acid on a reaction product in the presence of CMP and also found that formed CMP can be degraded enzymatically to thereby efficiently produce a sialic acid-containing sugar chain. The present inventors have further found that even a tetraantennary N-type sugar chain having four α2,6-linked sialic acid molecules, which has previously been difficult to synthesize, can be prepared at high yields by one-pot synthesis comprising the elongation reaction of a biantennary sugar chain used as a starting material without performing purification after each enzymatic reaction.

    摘要翻译： [待解决的问题]已知具有α2,3-或α2,6连接的唾液酸在其非还原性末端的糖链的重要性。 这些糖链化合物已经需要工业生产。 特别是通过控制唾液酸的连接模式（α2,6-键或α2,3-键），糖蛋白药物等的生产不可避免地需要通过量产生具有均匀结构的糖链。 特别地，在所有非还原性末端中的每一个具有唾液酸的三末端或四末端N-型复合糖链通常被认为难以化学合成。 没有报道披露这样的糖链是化学合成的。 此外，这些糖链也难以有效地制备酶。 [解决方案]本发明人在CMP存在下新发现唾液酸转移酶唾液酸转移酶在反应产物上的活性，并且发现形成的CMP可以酶促降解，从而有效地产生含唾液酸的糖链。 本发明人进一步发现，即使具有四个α2,6-连接的唾液酸分子的四末端N-型糖链，其先前难以合成，可以通过一锅合成以高收率制备，包括： 在每个酶反应后不用进行纯化的双天线糖链用作原料。

    N-acetylgalactosamine transferases and nucleic acids encoding the same
    37.
    发明授权
    N-acetylgalactosamine transferases and nucleic acids encoding the same 失效
    N-乙酰半乳糖胺转移酶和编码相同的核酸

    公开(公告)号：US08278037B2

    公开(公告)日：2012-10-02

    申请号：US12353385

    申请日：2009-01-14

    申请人： Hisashi Narimatsu ,  Masanori Gotoh ,  Takashi Sato

    发明人： Hisashi Narimatsu ,  Masanori Gotoh ,  Takashi Sato

    IPC分类号： C12N15/54 ,  C12N15/63 ,  C12Q1/68 ,  C12N9/10

    CPC分类号： C12N9/1051 ,  G01N33/57484

    摘要： An enzyme which transfers N-acetylgalactosamine to N-acetylglucosamine via a β1-4 linkage was isolated and the structure of its gene was explained. This led to the production of said enzyme or the like by genetic engineering techniques, the production of oligosaccharides using said enzyme, and the diagnosis of diseases on the basis of said gene or the like.The present invention uses a protein having the amino acid sequence shown in SEQ ID NO: 1, 3, 26 or 27 in the Sequence Listing or a variant of said amino acid sequence wherein one or more acids are substituted or deleted, or one or more acids are inserted or added and having the activity of transferring N-acetylgalactosamine (GalNAc) to N-acetylglucosamine serving as a substrate via a β1-4 linkage and nucleic acids encoding said protein.

    摘要翻译： 分离通过一个1-4键将N-乙酰半乳糖胺转移到N-乙酰氨基葡萄糖的酶，并解释其基因的结构。 这导致通过基因工程技术生产所述酶等，使用所述酶生产寡糖，以及基于所述基因等诊断疾病。 本发明使用具有序列表中SEQ ID NO：1,3,26或27所示氨基酸序列的蛋白质或其中一个或多个酸被取代或缺失的所述氨基酸序列的变体，或一个或多个 酸被插入或添加并且具有通过β-连接和编码所述蛋白质的核酸将N-乙酰半乳糖胺（GalNAc）转移到用作底物的N-乙酰葡糖胺的活性。

    Method of Excising Sugar Chain from Glycoprotein, Method of Mass Spectrometry of Sugar Chain, and Method of Mass Spectrometry of Glycoprotein
    38.
    发明申请
    Method of Excising Sugar Chain from Glycoprotein, Method of Mass Spectrometry of Sugar Chain, and Method of Mass Spectrometry of Glycoprotein 审中-公开
    从糖蛋白中消除糖链的方法，糖链质谱法和糖蛋白质谱法

    公开(公告)号：US20090057549A1

    公开(公告)日：2009-03-05

    申请号：US11918405

    申请日：2006-04-07

    申请人： Satoshi Kimura ,  Akihiko Kameyama ,  Hisashi Narimatsu

    发明人： Satoshi Kimura ,  Akihiko Kameyama ,  Hisashi Narimatsu

    IPC分类号： B01D59/44 ,  C12P19/00

    CPC分类号： G01N33/6848 ,  C12Q1/34 ,  G01N2333/924 ,  G01N2400/00

    摘要： The present invention provides a method of enzymatic reaction on a membrane by a sugar chain releasing enzyme, for an MALDI-TOF MS analysis of a glycoprotein that is solid-phased on a membrane is conducted directly on the membrane; a method of mass spectrometry of a sugar chain in which a sugar chain, for the MALDI-TOF MS analysis of a sugar chain excised from a glycoprotein that is solid-phased on a membrane is conducted directly on the membrane; and a method of mass spectrometry of a glycoprotein, for the MALDI-TOF MS analysis of a glycoprotein of a glycoprotein that is solid-phased on a membrane is conducted directly on the membrane. A method of excising a sugar chain to obtain excised sugar chains by excising the sugar chains by dispensing a sugar chain releasing enzyme solution on a glycoprotein that is solid-phased on a carrier, wherein the sugar chain releasing enzyme solution is a solution containing a sugar chain releasing enzyme in a reaction buffer solution containing a buffering agent consisting essentially of a volatile component. A method of mass spectrometry of a sugar chain and a method of mass spectrometry of a glycoprotein using the method of excising the sugar chain.

    摘要翻译： 本发明提供了通过糖链释放酶对膜进行酶反应的方法，对膜上固相化的糖蛋白的MALDI-TOF MS分析直接在膜上进行; 糖链的质谱法，其中直接在膜上进行糖链从用于在膜上固相的糖蛋白切除的糖链的MALDI-TOF MS分析; 并且对膜进行直接进行固相化的糖蛋白的糖蛋白的MALDI-TOF MS分析的糖蛋白的质谱法。 通过将糖链释放酶溶液分配在载体上的糖蛋白上切除糖链来切除糖链以获得切除的糖链的方法，其中糖链释放酶溶液是含有糖的溶液 在含有基本上由挥发性组分组成的缓冲剂的反应缓冲溶液中的链释放酶。 使用糖链的切割方法，糖链的质谱法和糖蛋白的质谱法。

    Glycosyltransferase, nucleic acid encoding the glycosyltransferase and method of testing canceration using the nucleic acid
    40.
    发明授权
    Glycosyltransferase, nucleic acid encoding the glycosyltransferase and method of testing canceration using the nucleic acid 失效
    糖基转移酶，编码糖基转移酶的核酸和使用核酸测试癌变的方法

    公开(公告)号：US07396666B2

    公开(公告)日：2008-07-08

    申请号：US10539834

    申请日：2003-12-26

    申请人： Hisashi Narimatsu ,  Takashi Kudo ,  Akira Togayachi ,  Toru Hiruma

    发明人： Hisashi Narimatsu ,  Takashi Kudo ,  Akira Togayachi ,  Toru Hiruma

    IPC分类号： C12N9/10 ,  C12N15/00 ,  C12P21/06 ,  C12P33/20 ,  C12Q1/68 ,  C07H21/04 ,  C07K1/00

    CPC分类号： C12N9/1051

    摘要： A tumor marker nucleic acid of the present invention is concerned with a nucleic acid hybridizing under stringent conditions to a nucleotide sequence described in SEQ ID NO: 1 or a complementary nucleotide sequence thereof. A method of testing canceration of the present invention is a method comprising diagnosing a biological sample as being cancerous when the transcription level of the nucleic acid in the biological sample significantly exceeds that in a normal biological sample as a control. The present invention also relates to a β1,3-N-acetyl-D-glucosaminyltransferase protein having an activity of transferring N-acetyl-D-glucosamine from a donor substrate to an acceptor substrate through β1,3-linkage.

    摘要翻译： 本发明的肿瘤标志物核酸涉及在严格条件下与SEQ ID NO：1所述核苷酸序列或其互补核苷酸序列杂交的核酸。 测定本发明的癌症的方法是当生物样品中的核酸的转录水平显着超过作为对照的正常生物样品中的生物样品时，将生物样品诊断为癌变的方法。 本发明还涉及具有通过β1,3键将N-乙酰基-D-葡糖胺从供体底物转移到受体底物的活性的β1,3-N-乙酰基-D-葡糖胺基转移酶蛋白。