Establishment and evaluation of in vitro galectin-3 inhibitor screening model
10.16438/j.0513-4870.2022-0342
- VernacularTitle:Galectin-3抑制剂体外筛选方法的建立及评价
- Author:
Chun-xiao MA
1
;
Xiao-wei XING
1
;
Shao-cong HOU
1
;
Shu-wang HE
2
;
Shi-qiang YAN
2
;
Ping-ping LI
1
Author Information
1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
2. Shandong DYNE Marine Biopharmaceutical Co., Ltd., Weihai 264333, China
- Publication Type:Research Article
- Keywords:
galectin-3;
glycosylation;
screening;
chronic inflammation;
fluorescein isothiocyanate
- From:
Acta Pharmaceutica Sinica
2023;58(1):156-161
- CountryChina
- Language:Chinese
-
Abstract:
Galectin-3 (Gal-3) belongs to the galectin family and is specific in binding β-galactoside. Through its C-terminal domain, Gal-3 binds to the galactoside group of the glycosylated insulin receptor (IR) and inhibits IR signaling pathway, which leads to the insulin resistance. Thus, Gal-3 is a potential therapeutic target for the treatment of insulin resistance and type 2 diabetes. Here we report a simple Gal-3 screening model based on the property that Gal-3 binds to the galactoside. We expressed and purified human Gal-3 in Escherichia coli (E.coli), and labeled it with fluorescein isothiocyanate (FITC) in vitro. After incubating FITC labeled Gal-3 (Gal-3-FITC) with PANC-1 cells, which express glycosylated membrane protein, PANC-1 cells started to show green fluorescent signal due to the Gal-3-FITC binding to the glycosylated membrane protein. Gal-3 inhibitor disrupts the binding of Gal-3-FITC and PANC1 cells, subsequently leads to the decrease of the fluorescent signal in PANC-1 cells. We can evaluate the inhibitory efficiency of Gal-3 inhibitors through measurement of the fluorescent signal. Further studies show this model is simple, stable, and repeatable with a Z' factor between 0.7 and 0.85. In sum, we have successfully established an in vitro high-throughput screening model for Gal-3 inhibitors.
