Záckova Markéta, Jezek Petr
Institute of Physiology, Academy of Sciences of the Czech Republic, Prague.
Biosci Rep. 2002 Feb;22(1):33-46. doi: 10.1023/a:1016009022186.
Reconstitution of novel mitochondrial uncoupling proteins, human UCP2 and UCP3, expressed in yeast, was performed to characterize fatty acid (FA)-induced H+ efflux in the resulted proteoliposomes. We now demonstrate for the first time that representatives of physiologically abundant long chain FAs, saturated or unsaturated, activate H+ translocation in UCP2- and UCP3-proteoliposomes. Efficiency of lauric, palmitic or linoleic acid was roughly the same, but oleic acid induced faster H+ uniport. We have confirmed that ATP and GTP inhibit such FA-induced H+ uniport mediated by UCP2 and UCP3. Coenzyme Q10 did not further significantly activate the observed H+ efflux. In conclusion, careful instant reconstitution yields intact functional recombinant proteins, UCP2 and UCP3, the activity of which is comparable with UCP1.
对在酵母中表达的新型线粒体解偶联蛋白——人UCP2和UCP3进行了重组，以表征脂肪酸（FA）诱导的质子外流在所得脂质体中的情况。我们现在首次证明，生理上丰富的长链脂肪酸（饱和或不饱和）的代表物可激活UCP2和UCP3脂质体中的质子转运。月桂酸、棕榈酸或亚油酸的效率大致相同，但油酸诱导的质子单向转运更快。我们已经证实，ATP和GTP可抑制由UCP2和UCP3介导的这种FA诱导的质子单向转运。辅酶Q10并没有进一步显著激活观察到的质子外流。总之，精心的即时重组可产生完整的功能性重组蛋白UCP2和UCP3，其活性与UCP1相当。
