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    • 3. 发明授权
    • Cdk5—specific inhibitory peptides
    • Cdk5特异性抑制肽
    • US06693166B1
    • 2004-02-17
    • US09289612
    • 1999-04-12
    • Jerry H. WangMingjie ZhangDamu Tang
    • Jerry H. WangMingjie ZhangDamu Tang
    • A61K3800
    • A61K38/005A61K38/1709
    • The activation of cyclin dependent kinase 5 (Cdk5) depends on the binding of its neuronal specific activator Nck5a. The minimal activation domain of Nck5a has been experimentally determine comprise acid residues 150 to 291 of the protein. It has been demonstrated that a 28 residue peptide encompassing amino acid residues Ala 146 to Asp 173 of Nck5a is capable of binding Cdk5 and hence resulting in the inhibition of its Kinase activity. Additionally, this peptide could also inhibit Cdk2 with a similar potency as it does to Cdk5. The direct competition experiments showed that the Nck5a inhibitory peptide does not compete with Nck5a for Cdk5 or cyclin A for Cdk2. Steady state kinetic analysis indicated that the Nck5a peptide acts as a non-competitive inhibitor of Cdk5/Nck5a complex with respect to its substrate. The structure of the peptide in solution as determined by the methods of circular dichroism and two-dimensional 1H NMR spectroscopy have been performed in order to understand the molecular basis of kinase inhibition by the peptide. A segment of the peptide, corresponding to amino acid resides Ser149 to Arg169 adopts an amphipathic alpha-helical structure in solution. Four Leu residues and one Phe residue clustered on the hydrophobic face of the helix, and this hydrophobic face is likely to be the contact area when the peptide binds and inhibits both Cdk5 and Cdk2. Mutational experiments have also indicated that the C-terminal end of the peptide contributes to the inhibition of Cdk5 and Cdk2.
    • 细胞周期蛋白依赖性激酶5(Cdk5)的激活取决于其神经元特异性活化剂Nck5a的结合。 Nck5a的最小活化结构域已经通过实验测定包含蛋白质的酸残基150至291。 已经证明,包含Nck5a的氨基酸残基Ala146至Asp173的28个残基肽能够结合Cdk5,因此导致其激酶活性的抑制。 另外,这种肽也可以以与Cdk5相同的效力抑制Cdk2。 直接竞争实验表明,Nck5a抑制肽与Cdk5或Cdk2的细胞周期蛋白A不同Nck5a竞争。 稳态动力学分析表明,Nck5a肽作为Cdk5 / Nck5a复合物相对于其底物的非竞争性抑制剂。 已经进行了通过圆二色性和二维1 H NMR光谱法测定的溶液中肽的结构,以了解肽激酶抑制的分子基础。 肽对应于氨基酸的片段Ser149至Arg169在溶液中采用两亲性α-螺旋结构。 四个Leu残基和一个Phe残基聚集在螺旋的疏水面上,当该肽结合并抑制Cdk5和Cdk2时,该疏水性面可能是接触面积。 突变实验也表明肽的C末端有助于抑制Cdk5和Cdk2。