title: Molecular structure, tissue distribution and functional characterization of interferon-γ-inducible lysosomal thiol reductase (GILT) gene in chicken (Gallus gallus).
abstract: Interferon-γ-inducible-lysosomal thiol reductase (GILT) plays a key role in the processing and presentation of MHC class II-restricted antigen (Ag) by catalyzing disulfide bond reduction, thus unfolding native protein Ag and facilitating subsequent cleavage by proteases. In this study, we reported the cloning of a GILT gene homologue from chicken (designated cGILT). The open reading frame (ORF) of cGILT consists of 762 bases, encoding a protein of 253 amino acids, with a putative molecular weight of 28kDa. The deduced protein possesses the typical structural feature of known GILT proteins, including an active-site motif, a GILT signature sequence, and 6 conserved cysteines. Genomic analysis revealed that cGILT gene, spanning a 1868bp fragment, contained seven exons interrupted by six introns. The result of real-time PCR showed that cGILT mRNA was expressed in a tissue-specific manner, while the cGILT mRNA expression was obviously up-regulated in spleen and PBMCs after stimulation with lipopolysaccharide (LPS). After expression as a soluble protein in Escherichia coli and purification by Ni-NTA affinity chromatography, cGILT was demonstrated to exhibit thiol reductase activity on IgG substrate.
authors.affiliation: Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Life Sciences College, Nanjing Normal University, Nanjing 210046, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Yang
authors.forename: Xiang
authors.initials: X
authors.lastname: Cao
authors.forename: Xuemei
authors.initials: X
authors.lastname: Ji
authors.forename: Hongzhen
authors.initials: H
authors.lastname: Liu
authors.forename: Haitao
authors.initials: H
authors.lastname: Wu
authors.forename: Wei
authors.initials: W
authors.lastname: Gu
authors.forename: Shuangquan
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000235:genetics*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000032:analysis / Q000737:chemistry / Q000235:genetics* / Q000502:physiology
chemical_list: D050862:Oxidoreductases Acting on Sulfur Group Donors