Molecular cloning of lamprey uncoupling protein and assessment of its uncoupling activity using a yeast heterologous expression system |
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Authors: | Chen Wang Guoxun Sun Kun Chen Zhiyuan Lv Shiming Peng Xueyuan Jiang Yang Xiang Chenyu Zhang |
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Institution: | 1. College of Fisheries, Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan, Hubei 430070, China;2. Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University, Auburn, AL 36849-5519, USA |
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Abstract: | We report the molecular cloning of a novel cDNA fragment from lamprey encoding a 313-amino acid protein that is highly homologous to human uncoupling proteins (UCP). We therefore named the protein lamprey UCP. This lamprey UCP, rat UCP1, human UCP2, and human mitochondrial oxoglutarate carrier were individually expressed in Saccharomyces cerevisiae and the recombinant yeast mitochondria were isolated and assayed for the state 4 respiration rate and proton leak. Only UCP1 showed a strong (3.6-fold increase of the ratio of mitochondrial state 4 respiration rate to FCCP-stimulated fully uncoupled respiration rate) and GDP-inhibitable uncoupling activity, while the uncoupling activities of both UCP2 and lamprey UCP were relatively weak (1.5-fold and 1.4-fold, respectively) and GDP-insensitive. The oxoglutarate carrier had no effect on the studied parameters. In conclusion, the lamprey UCP has a mild, unregulated uncoupling activity in the yeast system, which resembles UCP2, but not UCP1. |
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