Molecular and biological characterization of gamma-interferon-inducible lysosomal thiol reductase in silver carp (Hypophthalmichthys molitrix)

被引：6

作者：

Cao, Fang ^{[1
]}

Wu, Haitao ^{[2
]}

Lv, Tongtong ^{[1
]}

Yang, Yunqing ^{[1
]}

Li, Yue ^{[1
]}

Liu, Shuaimei ^{[1
]}

Hu, Lingling ^{[1
]}

Xu, Xixi ^{[1
]}

Ma, Lei ^{[1
]}

Zhang, Xinyi ^{[1
]}

Li, Jianfeng ^{[3
]}

Bi, Xiaolin ^{[4
]}

Gu, Wei ^{[4
]}

Zhang, Shuangquan ^{[1
]}

机构：

[1] Nanjing Normal Univ, Life Sci Coll, Jiangsu Prov Key Lab Mol & Med Biotechnol, Nanjing 210023, Jiangsu, Peoples R China

[2] Nanjing Univ Chinese Med, Sch Med & Life Sci, Nanjing 210023, Jiangsu, Peoples R China

[3] Hangzhou Normal Univ, Inst Aging Res, Sch Med, Hangzhou 311121, Zhejiang, Peoples R China

[4] Nanjing Univ Chinese Med, Sch Pharm, Nanjing 210023, Jiangsu, Peoples R China

来源：

FISH & SHELLFISH IMMUNOLOGY | 2018年 / 79卷

基金：

中国国家自然科学基金;

关键词：

Silver carp; GILT; Thio1 reductase activity; Immune responses; INNATE IMMUNE-RESPONSE; GILT-FREE MICE; EXPRESSION ANALYSIS; LPS CHALLENGE; CUTTING EDGE; GENE; CLONING; IDENTIFICATION; INDUCTION;

D O I：

10.1016/j.fsi.2018.04.064

中图分类号：

S9 [水产、渔业];

学科分类号：

0908 ;

摘要：

Gamma-interferon-inducible lysosomal thiol reductase (GILT) plays an important role in the processing of major histocompatibility complex (MHC) class II-restricted antigens by catalyzing disulfide bonds reduction. Herein, a GILT homolog (ScGILT) was identified from silver carp. Its open reading frame covers 771 base pairs, encoding a protein of 256 amino acids that possesses GILT signature sequence CQHGX(2)ECXZNX(4)C, active-site CXXC motif, and two potential N-linked glycosylation sites. The predicted tertiary structures of ScGILT and other GILTS were quite similar in shape and positional arrangement of the key motifs. ScGILT mRNA was constitutively expressed in all detected tissues, with high-level expression in fish immune organs, spleen and head kidney. After stimulation with lipopolysaccharide, the expression of ScGILT mRNA significantly increased in spleen and head kidney cells, and ScGILT protein translocated to late endosomes and lysosomes in HeLa cells. Recombinant ScGILT fused with a His(6) tag was expressed and purified, and could reduce the interchain disulfide bonds of IgG at pH 4.5. These results suggested that ScGILT was capable of catalyzing disulfide bonds reduction, and then might play an important role in the processing of MHC class II-restricted antigens in silver carp.

引用

页码：73 / 78

页数：6

共 27 条

[1] Enzymatic reduction of disulfide bonds in lysosomes: Characterization of a Gamma-interferon-inducible lysosomal thiol reductase (GILT) [J].