Ito Kikukatsu, Matsukawa Kazushige, Kato Yoshiaki
Cryobiosystem Research Center, Faculty of Agriculture, Iwate University, Morioka, Iwate 020-8550, Japan.
Biochem Biophys Res Commun. 2006 Oct 13;349(1):383-90. doi: 10.1016/j.bbrc.2006.08.058. Epub 2006 Aug 22.
Skunk cabbage, Symplocarpus foetidus, expresses two uncoupling proteins (UCPs), termed SfUCPA and SfUCPB, in the thermogenic organ spadix. SfUCPB exhibits unique structural features characterized by the absence of the putative fifth transmembrane domain (TM5) observed in SfUCPA, which is structurally similar to UCP1, and is abundantly expressed in the thermogenic spadix. Here, we conducted a series of comparative analyses of UCPs with six transmembrane domains, SfUCPA and rat UCP1, and TM5-deficient SfUCPB, using a heterologous yeast expression system. All UCPs were successfully expressed and targeted to the mitochondria, although the expression level of SfUCPB protein was approximately 10% of rat UCP1. The growth rate, mitochondrial membrane potential, and ATP content were significantly lower in cells expressing SfUCPB than in those expressing rat UCP1 and SfUCPA. These results suggest that SfUCPB, a novel TM5-deficient UCP, acts as an uncoupling protein in yeast cells.
臭菘(Symplocarpus foetidus)在产热器官佛焰花序中表达两种解偶联蛋白(UCPs),分别称为SfUCPA和SfUCPB。SfUCPB具有独特的结构特征,其特点是在SfUCPA中未观察到假定的第五跨膜结构域(TM5),SfUCPA在结构上与UCP1相似,且在产热佛焰花序中大量表达。在此,我们使用异源酵母表达系统对具有六个跨膜结构域的UCPs、SfUCPA和大鼠UCP1以及缺乏TM5的SfUCPB进行了一系列比较分析。所有UCPs均成功表达并定位于线粒体,尽管SfUCPB蛋白的表达水平约为大鼠UCP1的10%。表达SfUCPB的细胞的生长速率、线粒体膜电位和ATP含量显著低于表达大鼠UCP1和SfUCPA的细胞。这些结果表明,新型缺乏TM5的UCP——SfUCPB在酵母细胞中作为一种解偶联蛋白发挥作用。