The role of protein structure in the mitochondrial import pathway. Unfolding of mitochondrially bound precursors is required for membrane translocation.
In order to examine the influence of protein structure on the post-translational import of a protein into mitochondria, the carboxyl-terminal 129 residues of F1-ATPase beta-subunit precursor (511aa) have been replaced with 61 residues of yeast copper metallothionein. Import of the F1 beta-copper metallothionein (beta CuMT) hybrid into mitochondria was as efficient as that of the F1 beta precursor in the absence of copper. Addition of copper to mitochondrial import reactions, which had no significant effect on import of the F1 beta-subunit precursor, blocked import of the beta CuMT protein. This copper-dependent transport block for the beta CuMT precursor occurred after the precursor was bound to mitochondria. Expression of the beta CuMT protein in vivo revealed that beta CuMT would bind copper and allow growth of a copper-sensitive yeast host on an otherwise inhibitory level of the cation as long as it was localized in the cytoplasm. These data indicate that the binding of copper by beta CuMT renders it refractile for partial unfolding which is necessary for its translocation into mitochondria. These observations provide an alternative scheme for the selection of mutants defective in mitochondrial import.
Chen
W J
WJ
Department of Biochemistry, Southwestern Graduate School of Biomedical Sciences, University of Texas Health Science Center at Dallas 75235.
Douglas
M G
MG
D000595:Amino Acid Sequence; D001483:Base Sequence; D001693:Biological Transport, Active; D003001:Cloning, Molecular; D003300:Copper / Q000378:metabolism; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D013329:Structure-Activity Relationship
D011498:Protein Precursors; D021382:Protein Sorting Signals; C007121:copper thionein; D003300:Copper; D008668:Metallothionein