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首页> 外文期刊>Journal of Neuroscience Methods >A highly sensitive peptide substrate for detecting two Abeta-degrading enzymes: neprilysin and insulin-degrading enzyme.
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A highly sensitive peptide substrate for detecting two Abeta-degrading enzymes: neprilysin and insulin-degrading enzyme.

机译：一种高度敏感的肽底物，可检测两种Abeta降解酶：中性溶酶和胰岛素降解酶。

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摘要

Neprilysin has been singled out as the most promising candidate for use in the degradation of Abeta as a therapy for Alzheimer's disease. In this study, a quenched fluorogenic peptide substrate containing the first seven residues of the Abeta peptide plus a C-terminal Cysteine residue was synthesized to detect neprilysin activity. A fluorophore was attached to the C-terminal Cysteine and its fluorescence was quenched by a quencher linked to the N-terminus of the peptide. When this peptide substrate was degraded by an endopeptidase, fluorescence was produced and proved to be a sensitive detection system for endopeptidase activity. Our results showed that this assay system was extremely sensitive to neprilysin and insulin-degrading enzyme, but insensitive, or much less sensitive, to other Abeta-degrading enzymes. As low as 0.1 nM of neprilysin and 0.2 nM of insulin-degrading enzyme can be detected.
机译：脑啡肽酶已被选为最有希望的候选物，可用于降解Abeta作为阿尔茨海默氏病的疗法。在这项研究中，合成了包含Abeta肽的前七个残基和C端半胱氨酸残基的淬灭的荧光肽底物，以检测中性溶酶的活性。荧光团附着于C末端半胱氨酸，其荧光被连接至肽N端的淬灭剂淬灭。当该肽底物被内肽酶降解时，产生荧光，并被证明是内肽酶活性的灵敏检测系统。我们的结果表明，该测定系统对脑啡肽酶和胰岛素降解酶极为敏感，但对其他Abeta降解酶不敏感或敏感程度较低。可以检测到低至0.1 nM的脑啡肽酶和0.2 nM的胰岛素降解酶。

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