Co-expression of a mammalian accessory trafficking protein enables functional expression of the rat MCT1 monocarboxylate transporter in Saccharomyces cerevisiae

被引:20
|
作者
Makuc, J
Cappellaro, C
Boles, E
机构
[1] Univ Dusseldorf, Inst Mikrobiol, D-40225 Dusseldorf, Germany
[2] Univ Frankfurt, Inst Mikrobiol, D-60439 Frankfurt, Germany
关键词
Saccharomyces cerevisiae; heterologous expression; membrane transporter; monocarboxylate transport; pyruvate; CD147;
D O I
10.1016/j.femsyr.2004.06.003
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have developed a new heterologous expression system for monocarboxylate transporters. The system is based on a Saccharomyces cerevisiae pyk1 mae1 jen1 triple-deletion strain that is auxotrophic for pyruvate and deficient in monocarboxylate uptake. Growth of the yeast cells on ethanol medium supplemented with pyruvate or lactate was dependent on the expression of a suitable monocarboxylate transporter. We have used the system to characterize the functional significance of interactions between the rat MCT1 transporter and its ancillary protein CD147. CD147 was shown to improve trafficking of MCTI to the plasma membrane and its uptake activity. Our results demonstrate a new strategy for the production of properly folded and correctly targeted membrane proteins in a microbial expression system by co-expression of appropriate accessory proteins. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:795 / 801
页数:7
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