Proteomic analysis of NUS1 mutant R290C interaction proteins and their potential roles in Lennox-Gastaut syndrome
10.3760/cma.j.cn115354-20240830-00521
- VernacularTitle:NUS1突变体R290C互作蛋白组学分析及其在Lennox-Gastaut综合征中的可能作用
- Author:
Lizhi CHEN
1
;
Xiaoyan SHI
;
Nanxiang SHEN
;
Cuixia FAN
;
Zilong YE
;
Wenbin LI
Author Information
1. 广州医科大学附属第二医院广州医科大学神经科学研究所，神经致病基因和离子通道病教育部重点实验室，广州 510260
- Keywords:
Lennox-Gastaut syndrome;
NUS1;
Mutant;
Proteomics
- From:
Chinese Journal of Neuromedicine
2024;23(11):1113-1119
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the changes in interaction proteome of NUS1 mutant R290C and their relations with pathogenicity of Lennox Gastaut syndrome (LGS). Methods:The wild-type and mutant NUS1(R290C) plasmids were constructed and transfected into human embryonic kidney HEK293T cells; 48 h after that, NUS1 protein expression in HEK293T cells was detected by Western blotting. Co-immunoprecipitation, silver nitrate staining, and proteomic analysis were used to analyze the proteins interacted with wild-type or mutant NUS1 and identify the differential interacting proteins. Enrichment analyses of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed to annotate the molecular function and signaling pathways involved in the differential proteins. DisGeNet database was used to analyze the association between differential proteins and human diseases. Protein-protein interaction (PPI) was used to analyze the interaction network of NUS1 with protein folding regulatory proteins (RTN4 and DHDDS) and developmental epileptic encephalopathy related proteins.Results:(1) There was no significant difference in NUS1 protein expression between the wild-type and mutant NUS1 transfected HEK293T cells ( t=0.536, P=0.620). (2) Compared with that with wild-type NUS1 plasmid, number of proteins interacting with mutant NUS1 plasmid was significantly reduced in the transfected cells; 310 differential interacting proteins were screened in the mutant NUS1. (3) GO and KEGG enrichment analyses showed that the differential proteins were mainly involved in protein folding reaction and translation regulation. (4) DisGeNet association analysis showed that the two most relevant proteins in the differential interacting proteins were associated with frontotemporal dementia and developmental epileptic encephalopathy. (5) PPI analysis showed that NUS1 may be involved in occurrence of neurological diseases such as LGS by affecting protein folding signaling pathways. Conclusion:NUS1 mutant R290C alters its interacting protein lineage and mediates the development of LGS and other neurological diseases probably by regulating protein folding-related signaling.
