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Functional Expression of NADP-dependent Retinol Dehydrogenase/Reductase and Determination of Its Subcellular Localization
Ge-Fei Liu1, Dong-Yang Huang2*, Jing Du1,4, Ze-Shi Cui3, Jin-Dan Song1
1 Key Laboratory of Cell Biology of Ministry of Public Health of China, China Medical University, Shenyang 110001, China; 2 Medical School of Shantou University, Shantou 515031, China; 3Experimental Technology Center, Chin
Abstract: Human liver NADP-dependent retinol dehydrogenase/ reductase (NRDR) was expressed by baculovirus protein expression system and its biochemical activity was identified by the recombinant protein. The results indicated the NRDR was a NADP-dependent retinal reductase whose Km and Vmax for retinal reduction were (2.8±0.24) mM and (0.468±0.036) mmol/(min·mg)， respectively. Both of the wild type NRDR which has a C-terminal peroxisome targeting signal SRL， and mutant type NRDR whose C-terminal targeting signal was deleted expression vectors were constructed and transfected into HeLa cells. Immunofluorescence double staining was performed to determinate the subcellular localization of NRDR. The acquired images indicated that NRDR was targeted to peroxisome and this process was C-terminal SRL dependent. These data proved that NRDR was a NADP-dependent retinal reductase whose subcellular location was peroxisome.
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