Kinase activity of novel receptor interacting protein kinase 3 mutants / 上海交通大学学报(医学版)
Journal of Shanghai Jiaotong University(Medical Science)
;
(12): 856-860, 2019.
Article
in Chinese
| WPRIM
| ID: wpr-843376
ABSTRACT
Objective:
To explore the kinase activity of novel receptor interacting protein kinase 3 (RIPK3) mutants.Methods:
The four amino acids (Q84WDF87) of RIPK3 were mutated respectively and these mutants were co-transfected with mixed lineage kinase domain like pseudokinase (MLKL) into HEK293T cells. The auto-phosphorylation of these mutants at S232 and phosphorylation of MLKL at S345 were detected by Western blotting. The interaction between RIPK3 and MLKL was tested by co-immunoprecipitation. The oligomerization of MLKL was detected by non-reducing gel.Results:
The kinase activities of RIPK3ΔQ84, RIPK3ΔW85 and RIPK3ΔD86 were effectively decreased. Nevertheless, the kinase activities of RIPK3Q84A/RIPK3Q84E, RIPK3W85Y and RIPK3D86A/RIPK3D86Y did not change markedly. The auto-phosphorylation of RIPK3W85A at S232 was decreased without affecting phosphorylation and oligomerization of MLKL.Conclusion:
The amino acid site Q84, W85 or D86 plays a critical role in RIPK3 kinase activity. The kinase activity of RIPK3W85A is decreased, but it does not affect MLKL.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Journal of Shanghai Jiaotong University(Medical Science)
Year:
2019
Type:
Article
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