A synthetic signal peptide blocks import of precursor proteins destined for the mitochondrial inner membrane or matrix

• Published in: The Journal of biological chemistry 1985-12, Vol.260 (30), p.16045-16048
• Main Authors: Gillespie, L L, Argan, C, Taneja, A T, Hodges, R S, Freeman, K B, Shore, G C
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Intracellular Membranes - metabolism; Mitochondria, Liver - metabolism; Ornithine Carbamoyltransferase - genetics; Protein Precursors - genetics; Protein Processing, Post-Translational - drug effects; Protein Sorting Signals - chemical synthesis; Protein Sorting Signals - pharmacology; Rats; Ribonucleoproteins - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)36195-1

A peptide corresponding to amino acids 1-27 of preornithine carbamyltransferase (pOCT) has been chemically synthesized. When added to energized mitochondria in vitro, 20 microM of the peptide, designated pO(1-27), resulted in a collapse of the electrochemical potential across the mitochondrial inner membrane. This effect on transmembrane potential was not observed, however, when pO(1-27) was added to energized mitochondria under conditions that support in vitro import of precursor proteins (i.e. in the presence of reticulocyte lysate). The latter finding, therefore, made possible an examination of the ability of pO(1-27) to block import of homologous and heterologous proteins into the organelle. At 5-10 microM, pO(1-27) prevented import of pOCT in vitro; inhibition was overcome by increasing the concentration of pOCT. In contrast, pO(16-27), a peptide corresponding to amino acids 16-27 of pOCT and exhibiting a charge:mass ratio similar to pO(1-27) had no such inhibitory effect. pO(1-27) blocked import of other unrelated precursor proteins destined either for the mitochondrial matrix (pre-malate dehydrogenase and a hybrid protein containing the signal sequence of pre-carbamyl phosphate synthetase) or for the mitochondrial inner membrane (pre-thermogenin).