title: The mitochondrial targeting function of randomly generated peptide sequences correlates with predicted helical amphiphilicity.
abstract: A pool of oligonucleotides encoding a start methionine and nine random amino acids was inserted at the 5'-end of the gene for the yeast cytochrome oxidase subunit IV lacking its own mitochondrial targeting sequence. Approximately one-quarter of the randomly generated sequences targeted subunit IV to its correct intramitochondrial location in vivo. Sequence analysis of 89 randomly generated sequences showed that their efficiencies as mitochondrial targeting signals correlated with the potential to fold into an amphiphilic alpha-helix. Functional targeting sequences were enriched in arginine and isoleucine residues but contained few aspartate, glutamate, and proline residues. Nonfunctional sequences predicted to have significant helical amphiphilicity often had at least one acidic or multiple helix-breaking residues that would be expected to interfere with targeting functioning. These results support the hypothesis that the signal for targeting a protein into the mitochondrial matrix is usually a positively charged amphiphilic helix.
authors.affiliation: Biocenter, University of Basel, Switzerland.
authors.forename: B D
authors.initials: BD
authors.lastname: Lemire
authors.forename: C
authors.initials: C
authors.lastname: Fankhauser
authors.forename: A
authors.initials: A
authors.lastname: Baker
authors.forename: G
authors.initials: G
authors.lastname: Schatz
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003576:Electron Transport Complex IV / Q000235:genetics*; D015870:Gene Expression; D005800:Genes, Fungal; D005822:Genetic Vectors; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D010957:Plasmids; D011487:Protein Conformation; D011897:Random Allocation; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*
chemical_list: D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D003576:Electron Transport Complex IV