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KMID : 0545120140240040568
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Journal of Microbiology and Biotechnology
2014 Volume.24 No. 4 p.568 ~ p.576
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Construction and Characterization of an Enhanced GFP-Tagged TIM-1 Fusion Protein
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Jilin Qing
Haibing Xiao
Lin Zhao
Guifang Qin
Lihua Hu
Zhizhong Chen
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Abstract
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TIM-1 (also known as KIM-1 and HAVcr-1) is a type I transmembrane glycoprotein member of the TIM family that may play important roles in innate and adaptive immune responses. The overexpression of proteins associated with membrane proteins is a major obstacle to overcome in studies of membrane protein structures and functions. In this study, we successfully coupled the overexpression of the TIM-1 protein with a C-terminal enhanced green fluorescent protein (GFP) tag in Escherichia coli. To the best of our knowledge, this report is the first to describe the overexpression of human TIM-1 in E. coli. The purified TIM-1-EGFP fusion protein recognized and bound directly to apoptotic cells and did not to bind to viable cells. Furthermore, we confirmed that the interactions of TIM-1-EGFP with apoptotic cells were blocked by TIM-1-Fc fusion proteins. This fusion protein represents a readily obtainable source of biologically active TIM-1 that may prove useful in future studies of human TIM-1.
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KEYWORD
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HAVCR-1, fusion protein, apoptosis, E. coli, membrane protein, EGFP
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