title: Construction and characterization of bifunctional TIM-3-EGFP fusion proteins.
abstract: T cell immunoglobulin mucin-3 (TIM-3) is the first surface molecule to be found that specifically identifies Th1 cells in both mice and humans, and it negatively regulates Th1 responses. TIM-3 protein is a type I membrane protein. Overexpression of membrane proteins is a major problem to overcome in studies of membrane protein structure and function. In this study, a fusion between the gene encoding human TIM-3 and EGFP was successfully constructed and expressed in Escherichia coli. To our knowledge, this is the first time that human TIM-3 has been overexpressed in E. coli. We showed that the TIM-3-EGFP fusion protein mediates the recognition and binding of apoptotic cells. Furthermore, we demonstrated that the interactions of TIM-3-EGFP with apoptotic cells were blocked by TIM-3-Ig fusion proteins. Taken together, these results suggest that the human TIM-3 protein may act as a receptor for apoptotic cells.
authors.affiliation: Department of Laboratory, The People's Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, PR China.
authors.forename: Zhizhong
authors.initials: Z
authors.lastname: Chen
authors.forename: Jilin
authors.initials: J
authors.lastname: Qing
authors.forename: Guifang
authors.initials: G
authors.lastname: Qin
authors.forename: Lihua
authors.initials: L
authors.lastname: Hu
mesh_terms: D000818:Animals; D017209:Apoptosis; D016466:CHO Cells; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D056945:Hep G2 Cells; D000072597:Hepatitis A Virus Cellular Receptor 2; D006801:Humans; D020014:K562 Cells; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: C448908:HAVCR2 protein, human; D000072597:Hepatitis A Virus Cellular Receptor 2; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins