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    Two non-contiguous regions contribute to nidogen binding to a single EGF-like motif of the laminin .gamma.1 chain
    5.
    发明授权
    Two non-contiguous regions contribute to nidogen binding to a single EGF-like motif of the laminin .gamma.1 chain 失效
    两个不连续的区域有助于与层粘连蛋白γ1链的单个EGF样基序的结合

    公开(公告)号：US5493008A

    公开(公告)日：1996-02-20

    申请号：US288728

    申请日：1994-08-15

    Applicant: Jay W. Fox ,  Rupert Timpl

    Inventor： Jay W. Fox ,  Rupert Timpl

    IPC: C12N15/09 ,  A61K38/00 ,  A61P35/00 ,  A61P43/00 ,  C07K7/06 ,  C07K7/08 ,  C07K14/00 ,  C07K14/485 ,  C07K14/78 ,  C12P21/02 ,  C07K5/00

    CPC classification number: C07K14/78 ,  A61K38/00

    Abstract: High affinity binding of nidogen to laminin is mediated by an EGF-like repeat .gamma.1III4 of the mouse laminin .gamma.1 chain and has now been restricted to two short non-contiguous regions of its 56 residue sequence by use of synthetic peptides and recombinant mutants. Disulfide loop a,b of the repeat and a modified loop a,c could completely inhibit binding, with a 5,000-fold or 300-fold reduced affinity, respectively. Synthetic loops c and d lacked inhibitory activity. Some binding contribution of Try819 in loop c was, however, shown by mutation and side chain modification. Together with studies of loop chimeras, this indicated a distinct cooperativity between the two binding sites. The major binding site of loop a was localized to the heptapeptide NIDPNAV (position 798-804). A change of Asp800 to Asn or Ala803 to Val caused a strong reduction in binding activity, while only small effects were observed for the changes Pro801 to Gln and Ile799 to Val. The latter replacement corresponds to the single substitution found in the same region of the Drosophila laminin .gamma.1 chain. However, the changes Asn802 to Ser or Val804 to Ser, both known to exist in the laminin .gamma.2 chain, were deleterious mutations. This demonstrated conservation of binding structure in laminins of distantly related species, but not between homologous chains of laminin isoforms.

    Abstract translation: Nidogen与层粘连蛋白的高亲和力结合由小鼠层粘连蛋白γ1链的EGF样重复γ1III4介导，并且现在通过使用合成肽和重组突变体限制在其56个残基序列的两个短非连续区域。 二硫化物环a，b的重复和修饰的环a，c可分别完全抑制结合，减少亲和力为5000倍或300倍。 合成环c和d缺乏抑制活性。 然而，通过突变和侧链修饰显示Try819在环C中的一些结合贡献。 与循环嵌合体的研究一起，这表明两个结合位点之间具有明显的协同作用。 环a的主要结合位点定位于七肽NIDPNAV（798-804位）。 将Asp800转变成Asn或Ala803至Val引起结合活性的强烈降低，而对于Pro801至Gln和Ile799至Val的变化仅观察到微小的影响。 后一种替代物对应于在果蝇层粘连蛋白γ1链的相同区域中发现的单个取代。 然而，已知存在于层粘连蛋白γ2链中的Asn802至Ser或Val804至Ser的变化是有害突变。 这证明了在远处相关物种的层粘连蛋白中的结合结构的保守性，但不表现在层粘连蛋白同种型的同源链之间。

    Procedure for immunologic determination of procollagen peptide (type III) and procollagen peptide col 1 (type III) together and a process for preparing anti-procollagen peptide col 1 (type III) serum
    7.
    发明授权
    Procedure for immunologic determination of procollagen peptide (type III) and procollagen peptide col 1 (type III) together and a process for preparing anti-procollagen peptide col 1 (type III) serum 失效
    前胶原肽（III型）和前胶原肽1（III型）的免疫测定方法和制备抗原胶原肽col 1（III型）血清的方法

    公开(公告)号：US4504587A

    公开(公告)日：1985-03-12

    申请号：US474457

    申请日：1983-03-11

    Applicant: Rupert Timpl ,  Dietrich Brocks ,  Horst Neubauer ,  Helmut Strecker

    Inventor： Rupert Timpl ,  Dietrich Brocks ,  Horst Neubauer ,  Helmut Strecker

    IPC: G01N33/53 ,  A61K38/17 ,  A61K39/00 ,  A61K39/395 ,  G01N33/537 ,  G01N33/68 ,  G01N33/52 ,  G01N33/54 ,  G01N33/56

    CPC classification number: G01N33/537 ,  G01N33/6887 ,  Y10S436/80 ,  Y10S436/804 ,  Y10S436/811 ,  Y10S436/819 ,  Y10S436/824

    Abstract: The antigens procollagen peptide (type (III) and procollagen peptide col 1 (type III) can be determined together immunologically by either(a) reacting a specified amount in each case of labeled procollagen peptide (type III) or procollagen peptide col 1 (type III) and a highly specific antiserum containing antibodies having affinity for both the antigens mentioned together with a sample having an unknown content of procollagen peptide (type III) and/or procollagen peptide col 1 (type III), separating off the antigen-antibody complex formed and measuring the amount of labeling in the complex and/or in the supernatant, or(b) bringing a specified amount of the highly specific antiserum to reaction with a sample having an unknown content of procollagen peptide (type III) and/or procollagen peptide col 1 (type III), fixing the unreacted amount of the antibody to procollagen peptide (type III) or procollagen peptide col 1 (type III) bound to a support, and bringing to reaction with a labeled second antibody, and then determining the amount of bound or excess second antibody by measuring the labeling. An anti-procollagen peptide col 1 (type III) serum is preferred as the highly specific serum, and this is obtained by using procollagen peptide col 1 (type III) for immunizing experimental animals and obtaining their serum.

    Abstract translation: 抗原原胶原肽（类型（III）和前胶原肽肽1（III型））可以通过以下方式一起免疫测定：（a）在标记的前胶原肽（III型）或前胶原肽肽1（类型 III）和具有抗体的高度特异性抗血清，所述抗体与所述抗原一起提供与具有未知含量的原胶原肽（III型）和/或前胶原肽第1号（III型）的样品的亲和力，分离出抗原 - 抗体复合物 形成和测量复合物和/或上清液中的标记量，或（b）使特定量的高度特异性抗血清与具有未知含量的前胶原肽（III型）和/或前胶原的样品反应 肽柱1（III型），将未反应量的抗体固定到与载体结合的前胶原肽（III型）或前胶原肽肽1（III型）上，并与标记物反应 然后通过测量标记来确定结合或过量的第二抗体的量。 优选抗原胶原肽col 1（III型）血清作为高度特异性血清，并且通过使用前胶原肽肽1（III型）免疫实验动物并获得其血清而获得。

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