Identification and activity analysis of ubiquitin ligase TP12446 gene from Trichinella spiralis
10.16303/j.cnki.1005-4545.2025.05.18
- VernacularTitle:旋毛虫泛素连接酶TP12446基因的活性鉴定及分析
- Author:
Shuyan ZHANG
1
;
Zijian DONG
1
;
Jianda PANG
1
;
Saining WANG
1
;
Qianqian DANG
1
;
Fengying YIN
1
;
Xiaolei LIU
1
;
Xuelin WANG
1
Author Information
1. 吉林大学动物医学学院人兽共患病研究所/人畜共患传染病重症诊治全国重点实验室/人兽共患病研究教育部重点实验室,吉林长春 130062
- Publication Type:Journal Article
- Keywords:
Trichinella spiralis;
ubiquitin ligase;
bioinformatics;
identification of activity
- From:
Chinese Journal of Veterinary Science
2025;45(5):1017-1025
- CountryChina
- Language:Chinese
-
Abstract:
Based on the previous transcriptomic experimental data of Trichinella spiralis(T.spira-lis)in this study,the larval stage specific gene TP12446 was screened and its identity in the ubiq-uitin ligase RNF family was predicted.In the study,bioinformatics methods were used to analyze its physicochemical properties and its activity to lay the foundation for further exploring the func-tion of TP12446 gene.The physicochemical properties and protein structure of TP12446 protein were predicted by bioinformatics.Its ubiquitin ligase activity was also verified by ubiquitination re-actions in vitro.The expression characteristics of TP12446 protein in different stage of T.spiralis infection were analyzed by qPCR and Western blot.Bioinformatics analysis showed that TP12446 protein was composed of 453 amino acids and its molecular weight was 51.48 kDa.The protein had a transmembrane structure and contained signal peptides.The results indicated that it was a secre-tory protein and mainly located in the cytoplasmic membrane.The protein structure analysis re-vealed that the protein contained RING and PA domain,its secondary structure was mainly com-posed of α-helix and irregular crimp and there were 10 B cell epitopes on TP12446 protein.The prediction of glycosylation and phosphorylation sites indicated that TP12446 protein contained 38 potential phosphorylation sites.Results of PPI interaction protein prediction showed that TP12446 protein had strong interaction with Usp8,Tmem37,Otub1,Otub2,Ubox5 and CD151.The results of qPCR and Western blot showed that TP12446 gene expression was the highest in the larva stage of T.spiralis,the activity of ubiquitin ligase was verified by ubiquitination reaction in vitro.TP12446 protein was a secretory hydrophobic protein with E3 ubiquitin ligase activity,which was involved in regulating cell cycle and apoptosis.
