A screen assay of ?-secretase inhibitors in vitro
- VernacularTitle:?分泌酶抑制剂体外筛选模型的构建与验证
- Author:
Bin ZHAO
;
Hongjun YANG
;
Jing ZHAO
- Publication Type:Journal Article
- Keywords:
?-secretase;
ELISA;
screen assay;
Alzheimers
- From:
Chinese Pharmacological Bulletin
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To construct a screen assay of ?-secretase inhibitors.Methods A human ?-secretase cDNA was cloned by RT-PCR and then reconstructed in an expression vector.The recombinant ?-secretase fusion protein was expressed and purified from Escherichia coli.A screen assay of ?-secretase inhibitors was constructed in vitro using a recombinant A ?-fusion protein as substrate.The enzyme reaction was detected by SDS-PAGE and an ELISA method using metal chelating plates to catch the substrate.Results A human ?-secretaae cDNA was cloned and expressed in escherichia coli.The ?-secretase enzyme reaction can be detected by two kinds of method: SDS-PAGE and ELISA.The IC_(50) for an inhibitor statVal determined by this method was in good agreement with those reported in the literature.Seventy seven species of Chinese traditional medical plants were screened using this method and seventeen species show weak inhibitory activity.Conclusion A screen assay of ?-secretase inhibitors was successfully constructed and can be used to screen natural products.
