Polypeptide capable of inhibiting beta-secretase digestion effect and application thereof
A technology for preparations and active ingredients, which is applied to polypeptides inhibiting β-secretase enzymatic cleavage and their application fields, and can solve problems such as allergy, large antibody molecules, and difficulty.
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Embodiment 1
[0043] Example 1, the acquisition of polypeptide YS1 and its derivatives
[0044] 1. Obtaining the polypeptide YS1
[0045]According to the pathogenesis of Alzheimer's disease AD, the fundamental way to treat AD should be to reduce the production of β-amyloid (Aβ), inhibit the aggregation of Aβ or accelerate the clearance of Aβ. Using phage display technology, using the β-secretase cleavage site on APP and the partial amino-terminal sequence of Aβ as the screening substrate, 1×10 8 The heptapeptides were panned for four rounds, and a number of peptides that could bind to the β-secretase cleavage site and Aβ42 on APP were screened out by phage ELISA. The cleavage site of β-secretase and the binding characteristics of Aβ42 and the influence on Aβ aggregation and cytotoxicity finally select the polypeptide YS1 through the combination of peptide chains.
[0046] The amino acid sequence of YS1 is: Pro-Gln-Val-Gly-His-Leu (Sequence 1, PQVGHL).
[0047] Peptide YS1 is a hexapeptid...
Embodiment 2
[0055] Embodiment 2, the application of polypeptide YS1
[0056] 1. The specific binding of polypeptide YS1 and its derivatives to the β-secretase cleavage site on APP and Aβ42
[0057] 1. The specific binding of polypeptide YS1 to the β-secretase cleavage site on APP and Aβ42
[0058] The β-secretase cleavage site polypeptide (Glu-Val-Lys-Met-Asp-Glu-Phe) (synthesized by Shanghai Gil Polypeptide Co.), Aβ42 (American Peptide Co., catalog No. 62-0-80B) and negative control polypeptide (Lys-Thr-Asp-Met-Lys-His-Met-Ala-Gly-Ala-Ala-Ala-Ala-Gly-Ala-Val-Val-Gly-Gly -Leu-Gly) was coated with 1 μg / well on a 96-well aminolation microtiter plate, overnight at 4°C. Block the blank site on the microtiter plate that is not bound to the polypeptide with BSA, then add YS1 (1 μg / well) obtained in Example 1 with 6 histidines attached as tags, and act for 2 hours at room temperature (25°C). After washing, an HRP-coupled antibody capable of binding to the histidine tag (Beijing Boaosen Biotec...
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