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    Heregulin variants
    1.
    发明授权
    Heregulin variants 失效
    特殊蛋白变体

    公开(公告)号：US06387638B1

    公开(公告)日：2002-05-14

    申请号：US09101544

    申请日：1998-07-17

    申请人： Marcus D. Ballinger ,  Jennifer T. Jones ,  Wayne J. Fairbrother ,  Mark X. Sliwkowski ,  James A. Wells

    发明人： Marcus D. Ballinger ,  Jennifer T. Jones ,  Wayne J. Fairbrother ,  Mark X. Sliwkowski ,  James A. Wells

    IPC分类号： G01N33566

    CPC分类号： G01N33/74 ,  A61K38/00 ,  C07K14/4756 ,  G01N2333/485

    摘要： The present invention provides heregulin variants that are capable of binding an ErbB receptor. Included in the invention are variants of human heregulins, and, in particular, variants of human heregulin-&bgr;1 having enhanced affinity for the ErbB-3 and ErbB-4 receptors. These variants include at least one amino acid substitution and can include further modifications. The invention also provides nucleic acid molecules encoding heregulin variants and related vectors, host cells, pharmaceutical compositions, and methods.

    摘要翻译： 本发明提供能够结合ErbB受体的调节蛋白变体。 本发明中包括本发明的变体，尤其是具有对ErbB-3和ErbB-4受体具有增强亲和性的人类调节蛋白-β1的变体。 这些变体包括至少一个氨基酸取代并且可以包括进一步的修饰。 本发明还提供了编码本领域变体和相关载体的核酸分子，宿主细胞，药物组合物和方法。

    Heregulin variants
    3.
    发明授权
    Heregulin variants 失效
    特殊蛋白变体

    公开(公告)号：US6136558A

    公开(公告)日：2000-10-24

    申请号：US20880

    申请日：1998-02-09

    申请人： Marcus D. Ballinger ,  Jennifer T. Jones ,  Wayne J. Fairbrother ,  Mark X. Sliwkowski ,  James A. Wells

    发明人： Marcus D. Ballinger ,  Jennifer T. Jones ,  Wayne J. Fairbrother ,  Mark X. Sliwkowski ,  James A. Wells

    IPC分类号： C07K14/575 ,  C12P21/06

    CPC分类号： C07K14/57509

    摘要： The present invention provides heregulin variants that are capable of binding an ErbB receptor. Included in the invention are variants of human heregulins, and, in particular, variants of human heregulin-.beta.1 having enhanced affinity for the ErbB-3 and ErbB-4 receptors. These variants include at least one amino acid substitution and can include further modifications. The invention also provides nucleic acid molecules encoding heregulin variants and related vectors, host cells, pharmaceutical compositions, and methods.

    摘要翻译： 本发明提供能够结合ErbB受体的调节蛋白变体。 本发明中包括本发明的变体，尤其是具有对ErbB-3和ErbB-4受体具有增强亲和力的人类胰岛素β1的变体。 这些变体包括至少一个氨基酸取代并且可以包括进一步的修饰。 本发明还提供了编码本领域变体和相关载体的核酸分子，宿主细胞，药物组合物和方法。

    Subtilisin variants capable of cleaving substrates containing dibasic residues
    4.
    发明授权
    Subtilisin variants capable of cleaving substrates containing dibasic residues 失效
    能够切割含有二碱基残基的底物的枯草杆菌蛋白酶变体

    公开(公告)号：US5780285A

    公开(公告)日：1998-07-14

    申请号：US398028

    申请日：1995-03-03

    申请人： Marcus D. Ballinger ,  James A. Wells

    发明人： Marcus D. Ballinger ,  James A. Wells

    IPC分类号： C12N9/54 ,  C12N9/64 ,  C12N15/57 ,  C12N15/62 ,  C12P21/06 ,  C12N9/56 ,  C12N15/75

    CPC分类号： C12N9/6454 ,  C12N15/62 ,  C12N9/54 ,  C12P21/06 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/75

    摘要： The bacterial serine protease, subtilisin BPN', has been mutated so that it will efficiently and selectively cleave substrates containing dibasic residues. A combination mutant, where Asn 62 was changed to Asp and Gly 166 was changed to Asp (N62D/G166D), had a larger than additive shift in specificity toward dibasic substrates. Suitable substrates of the variant subtilisin were revealed by sorting a library of phage particles (substrate phage) containing five contiguous randomized residues. This method identified a particularly good substrate, Asn-Leu-Met-Arg-Lys-, that was selectively cleaved in the context of a fusion protein by the N62D/G166D subtilisin variant. Accordingly, this variant subtilisin may be useful for cleaving fusion proteins with dibasic substrate linkers and processing hormones or other proteins (in vitro or in vivo) that contain dibasic cleavage sites.

    摘要翻译： 细菌丝氨酸蛋白酶枯草杆菌蛋白酶BPN'已经被突变，使得其将有效地和选择性地裂解含有二元残基的底物。 将Asn 62改变为Asp和Gly 166的组合突变体改变为Asp（N62D / G166D），其特异性对于二元底物的添加偏移大。 通过分选含有五个连续随机残留物的噬菌体颗粒（底物噬菌体）文库来揭示变体枯草杆菌蛋白酶的合适底物。 该方法鉴定了特异性良好的底物Asn-Leu-Met-Arg-Lys-，其被N62D / G166D枯草杆菌蛋白酶变体在融合蛋白的上下文中选择性地切割。 因此，这种变体枯草杆菌蛋白酶可用于用含有二碱基切割位点的二碱基底物接头和加工激素或其他蛋白质（体外或体内）切割融合蛋白。

    Subtilisin variants capable of cleaving substrates containing basic residues
    5.
    发明授权
    Subtilisin variants capable of cleaving substrates containing basic residues 失效
    能够裂解含有碱性残基的底物的枯草杆菌蛋白酶变体

    公开(公告)号：US5837516A

    公开(公告)日：1998-11-17

    申请号：US504265

    申请日：1995-07-19

    申请人： Marcus D. Ballinger ,  James A. Wells

    发明人： Marcus D. Ballinger ,  James A. Wells

    IPC分类号： C12N9/54 ,  C12N9/64 ,  C12N15/57 ,  C12N15/62 ,  C12P21/06 ,  C12N15/75

    CPC分类号： C12N9/6454 ,  C12N15/62 ,  C12N9/54 ,  C12P21/06 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/75

    摘要： The bacterial serine protease, subtilisin BPN', has been mutated so that it will efficiently and selectively cleave substrates containing basic residues. Combination mutants, where Asn 62 was changed to Asp, Gly 166 was changed to Asp (N62D/G166D), and optionally Tyr 104 was changed to Asp had a larger than additive shift in specificity toward substrates containing basic residues. Suitable substrates of the variant subtilisin were revealed by sorting a library of phage particles (substrate phage) containing five contiguous randomized residues. This method identified a particularly good substrate, Asn-Leu-Met-Arg-Lys- (SEQ ID NO: 35), that was selectively cleaved in the context of a fusion protein by the N62D/G166D subtilisin variant. A particularly good substrate for N62D/G166D/Y104D would be Asn-Arg-Met-Arg-Lys- (SEQ ID NO: 76). Accordingly, these variant subtilisin are useful for cleaving fusion proteins with basic substrate linkers and processing hormones or other proteins (in vitro or in vivo) that contain basic cleavage sites.

    摘要翻译： 细菌丝氨酸蛋白酶枯草杆菌蛋白酶BPN'已被突变，从而能有效地选择性地切割含有碱性残基的底物。 将Asn 62改变为Asp的组合突变体，将Gly 166改变为Asp（N62D / G166D），并且任选地将Tyr 104改变为Asp，对于含有碱性残基的底物的特异性大于添加位移。 通过分选含有五个连续随机残留物的噬菌体颗粒（底物噬菌体）文库来揭示变体枯草杆菌蛋白酶的合适底物。 该方法鉴定了通过N62D / G166D枯草杆菌蛋白酶变体在融合蛋白的上下文中选择性切割的特别良好的底物Asn-Leu-Met-Arg-Lys-（SEQ ID NO：35）。 N62D / G166D / Y104D特别好的底物是Asn-Arg-Met-Arg-Lys-（SEQ ID NO：76）。 因此，这些变体枯草杆菌蛋白酶可用于用碱性底物接头和加工激素或其它含有碱基切割位点的蛋白质（体外或体内）切割融合蛋白。