Journal of Shanghai Jiao Tong University (Medical Science) >

2019 , Vol. 39 >Issue 8: 856

DOI: https://doi.org/10.3969/j.issn.1674-8115.2019.08.008

Original article (Basic research)

Kinase activity of novel receptor interacting protein kinase 3 mutants

  • ZHANG Yue1 ,
  • ZHANG Hai-wei2 ,
  • ZHANG Hai-bing2 ,
  • LUO Yan 1
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  • 1. Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China; 2. Key Laboratory of Nutrition and Metabolism, Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

Online published: 2019-09-23

Supported by

Foundation of Science and Technology Commission of Shanghai Municipality, 16ZR1421100

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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 detectedWestern blotting. The interaction between RIPK3 and MLKL was testedco-immunoprecipitation. The oligomerization of MLKL was detectednon-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.

Key words： receptor interacting protein kinase 3 (RIPK3); mixed lineage kinase domain like pseudokinase (MLKL); kinase activity; necroptosis

Cite this article

ZHANG Yue1 , ZHANG Hai-wei2 , ZHANG Hai-bing2 , LUO Yan 1 . Kinase activity of novel receptor interacting protein kinase 3 mutants[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2019 , 39(8) : 856 . DOI: 10.3969/j.issn.1674-8115.2019.08.008

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