Functional reconstitution of rat uncoupling protein following its high level expression in yeast.

摘要: Small mammals, including human infants, rely on nonshivering thermogenesis for a substantial portion of their body heat during exposure to cold. This is mediated in large part by the uncoupling protein, which found exclusively within inner membrane brown adipose tissue mitochondria. The sole function protein provide regulated transport pathway electrophoretic back-flux H+ ions into mitochondrial matrix, thereby dissipating protonmotive force and producing heat. Thus, unique with respect both its physiological role expression. We have now achieved high level expression rat yeast, import yeast mitochondria at levels, 70-100 micrograms/mg similar those observed from cold-adapted rats. When expressed was purified reconstituted liposomes, proteoliposomes exhibited GDP-sensitive proton chloride uniports that were inhibited GDP Ki values obtained native protein. Moreover, molecular activities Cl- indistinguishable availability unlimited amounts functional, will permit application site-directed mutagenesis many intriguing aspects structure function.