Effect of Amino Acid Motifs in Integrin β3 Cytoplasmic Tail on αⅡbβ3-Mediated Cell function in 293T cell models.
10.7534/j.issn.1009-2137.2019.01.037
- Author:
Dong-Ya LI
1
;
Jian-Hua MAO
2
;
Wei ZHANG
2
;
Xin-Jie CHEN
2
;
Bing XIAO
2
;
Zheng RUAN
2
;
Yun WANG
1
;
Guo-Xiong JIANG
1
;
Xiao-Feng SHI
1
;
Xiao-Dong XI
3
Author Information
1. Department of Hematology, Affiliated Hospital of Jiangsu University, Zhenjiang, 212100 Jiangsu Province, China.
2. State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
3. Department of Hematology, Affiliated Hospital of Jiangsu University, Zhenjiang, 212100 Jiangsu Province, China.E-mail: xixiaodong@shsmu.edu.cn.
- Publication Type:Journal Article
- MeSH:
Amino Acid Motifs;
Animals;
CHO Cells;
Cell Adhesion;
Cricetinae;
Cricetulus;
HEK293 Cells;
Humans;
Integrin beta3;
genetics;
metabolism;
Platelet Glycoprotein GPIIb-IIIa Complex;
genetics;
metabolism;
Signal Transduction
- From:
Journal of Experimental Hematology
2019;27(1):227-232
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To establish 293T cell lines stably expressing Calpain-cleavage related α3 cytoplasmic tail mutants, and to explore the effect of amino acid motifs in integrin β3 cytoplasmic tail on αⅡbβ3-mediated cell function.
METHODS:293T cell lines stably co-expressing human wild type integrin αⅡb and full length β3 or mutant β3, including β3-ΔNITY (β3 cytoplasmic tail NITY motif deleted), β3-Δ754 (β3 cytoplasmic tail TNITYRGT motif deleted) and β3-Δ759 (β3 cytoplasmic tail RGT motif deleted) were established. Spreading and adhesion of these stable cell lines on immobilized fibrinogen were tested.
RESULTS:293T-αⅡbβ3ΔNITY, 293T-αⅡbβ3Δ754, 293T-αⅡbβ3Δ759 and 293T-αⅡbβ3 cell lines were successfully established. Compared with the 293T cells, 293T-αⅡbβ3 cells which expressed full β3, possessed well adhesion and spread ability on immobilized fibrinogen, suggesting it can be as a surrogate for platelet. Compared with 293T-αⅡbβ3 cells, the 293T-αⅡbβ3ΔNITY cells showed a partial impairment of adhesion and spreadability on immobilized fibrinogen. while the 293T-αⅡbβ3Δ754 cells and 293T-αⅡbβ3Δ759 cells failed to adhere or spread on immobilized fibrinogen.
CONCLUSION:To the cell spreading function mediated by integrin β3, RGT motif is vital, while NITY can be dispensable. These established 293T cell lines stably expressing different β3 mutants provide a solid basis for a further analysis of mass spectrometry.
