The role and mechanism of chaperones Calnexin/Calreticulin in which ALLN selectively rescues the trafficking defective of HERG-A561V mutation
| Autor: | Ying Wang, Tingting Shen, Jianqing Zhou, Junbo Zhou, Ningsheng Liu, Kenan Lou, Jiangfang Lian, Peiliang Fang, Hai Qian, Zemin Cen |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
ERG1 Potassium Channel congenital hereditary and neonatal diseases and abnormalities Patch-Clamp Techniques Calnexin Leupeptins Immunoprecipitation ALLN hERG Biophysics 030204 cardiovascular system & hematology Biochemistry 03 medical and health sciences 0302 clinical medicine chaperones Calnexin/Calreticulin Western blot Mutant protein medicine Humans HERG-A561V mutation cardiovascular diseases Molecular Biology Research Articles biology medicine.diagnostic_test Chemistry Cell Biology Propranolol Transport protein Cell biology Long QT Syndrome Protein Transport HEK293 Cells 030104 developmental biology Chaperone (protein) Mutation biology.protein Calreticulin Research Article Molecular Chaperones |
| Zdroj: | Bioscience Reports |
| ISSN: | 1573-4935 0144-8463 |
| DOI: | 10.1042/bsr20171269 |
| Popis: | Long QT (LQT) type 2 (LQT2) is caused by HERG mutation. L539fs/47 encodes a truncated protein, and its mechanisms in HERG mutation are unknown. HERG mutation plasmids were overexpressed in HEK293T cells, respectively, followed by analyzing lysates with Western blot. Transfected HEK293T cells were treated with or without N-acetyl-l-leucyl-l-leucyl-l-norleucinal (ALLN) and Propranolol (Prop) at 24 or 48 h. HERG-WT, HERG-A561V, WT/A561V, HERG-L539fs/47, WT/L539fs/47, and Calnexin (CNX)/Calreticulin (CRT) protein expression and their interactions were detected by Western blot and immunoprecipitation. Each group with HERG currents (Ikr) were detected by Patch-clamp technique. Treated with ALLN, the expression of mature HERG protein and the CNX/CRT protein increased. The interaction of HERG-A561V and WT/A561V protein with the chaperone CNX/CRT increased significantly. The maximum peak currents and tail currents density increased by 70% and 73%, respectively, while maximal peak currents density (24%) and tail currents density (19%) were slight increased in WT-HERG cells. Treated with Prop, the expression and interaction of mature HERG and chaperones CNX/CRT had no difference in each group. The maximal currents and tail currents density were slight increased. CNX/CRT might play a crucial role in the trafficking-deficient process and degradation of HERG-A561V mutant protein, however they had no effect on L539fs/47 HERG due to protein transport deletion. ALLN can restore HERG-A561V mutant protein trafficking process and rescue the dominant negative suppression of WT-HERG. |
| Databáze: | OpenAIRE |
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