The adenine nucleotide translocase mediates the exchange of ADP and ATP across the inner mitochondrial membrane. Therefore, proper function of ANT is essential for the transfer of ATP synthesized in mitochondria to the cytoplasm. Most eukaryotes from yeast to humans have multiple ANT isoforms. Unicellular organisms utilize different ANT isoforms depending on the availability of external nutrients and aeration. Multicellular organisms, express different ANT isoforms in a tissue-specific manner that are apparently adapted to the unique metabolic demands of the various tissues. Recently, we and others identified a novel member of the ANT family, ANT4 in both humans and mice. ANT4 is evolutionarily conserved in mammals and Pirenperone exclusively expressed in male germ cells of adult animals. Although the previous gene knock-out study in mouse revealed that ANT4 was essential for the process of male germ cell meiosis in mice, neither the function of ANT4 in male germ cells nor the reason why ANT4 exists only within a limited spectrum of species is known. The human ANT4 gene is predicted to encode a 315 amino acid protein. The protein contains the characteristic amino acid sequence shared by all known ADP/ATP carriers. hANT4 was demonstrated to possesses bona fide ADP/ ATP transport upon reconstitution and assay of recombinant protein from E. coli into proteo-liposomes. The reported kinetics of hANT4 were distinct from previously reported kinetics of other somatic hANTs, with comparatively lower affinity for adenine nucleotides and a higher Vmax. Unfortunately, the kinetics of ADP/ATP transport through hANT4 and somatic hANTs were not compared under comparable experimental conditions. Therefore, it is important to evaluate the differing biochemical characteristics of hANT4 and the somatic hANTs to elucidate the functional role of hANT4. In order to determine the biochemical properties of the hANT4, we have chosen to heterologously express each hANT isoform in yeast and analyze their biochemical properties in PF-05020182 parallel. Baker��s yeast, Saccharomyces cerevisiae contains three paralogous genes encoding ADP/ATP carriers: AAC1, AAC2 and AAC3. Yeast AACs have been extensively studied, taking advantage of the myriad molecular and genetic tools available in this organism.