The precursor of mitochondrial aspartate aminotransferase is imported into mitochondria faster than the homologous cytosolic isoenzyme with the same presequence attached

Abstract

Mitochondrial and cytosolic aspartate aminotransferase (AspAT) are homologous proteins with identically folded polypeptide chains. The cDNAs of the two isoenzymes of chicken were used to express the following proteins in yeast: the precursor of mitochondrial AspAT, mature mitochondrial AspAT, and two chimeric proteins in one of which (pc) the presequence of the precursor was attached to the entire cytosolic isoenzyme and in the other one (pmc) the N-terminal segment (amino acid residues -22 to 23) of the precursor was linked to the slightly truncated cytosolic isoenzyme (residues 34 to 412). All presequence containing proteins were imported into the mitochondria and processed to the mature form whereas mature mitochondrial AspAT remained in the cytosol. The rate of import of the authentic precursor was four times faster than that of the chimeric proteins pc and pmc, t1/2 for importation at 29 degrees C being 3, 13 and 14 min, respectively. Apparently, the mature moiety of the precursor of mitochondrial AspAT promotes importation.