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. 1989 Nov;86(21):8432–8436. doi: 10.1073/pnas.86.21.8432

Translocation of proteins across the mitochondrial inner membrane, but not into the outer membrane, requires nucleoside triphosphates in the matrix.

S T Hwang 1, G Schatz 1
PMCID: PMC298296  PMID: 2554316

Abstract

Work in several laboratories has established that import of proteins across the mitochondrial inner membrane requires an electrochemical potential across that membrane and cleavage of nucleoside triphosphate. We now show that nucleoside triphosphate must be present inside the inner membrane. In contrast, the potential-independent insertion of porin into the outer membrane requires ATP only outside the inner membrane.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chen W. J., Douglas M. G. Phosphodiester bond cleavage outside mitochondria is required for the completion of protein import into the mitochondrial matrix. Cell. 1987 Jun 5;49(5):651–658. doi: 10.1016/0092-8674(87)90541-1. [DOI] [PubMed] [Google Scholar]
  2. Cheng M. Y., Hartl F. U., Martin J., Pollock R. A., Kalousek F., Neupert W., Hallberg E. M., Hallberg R. L., Horwich A. L. Mitochondrial heat-shock protein hsp60 is essential for assembly of proteins imported into yeast mitochondria. Nature. 1989 Feb 16;337(6208):620–625. doi: 10.1038/337620a0. [DOI] [PubMed] [Google Scholar]
  3. Daum G., Böhni P. C., Schatz G. Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J Biol Chem. 1982 Nov 10;257(21):13028–13033. [PubMed] [Google Scholar]
  4. Deshaies R. J., Koch B. D., Werner-Washburne M., Craig E. A., Schekman R. A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides. Nature. 1988 Apr 28;332(6167):800–805. doi: 10.1038/332800a0. [DOI] [PubMed] [Google Scholar]
  5. Eilers M., Hwang S., Schatz G. Unfolding and refolding of a purified precursor protein during import into isolated mitochondria. EMBO J. 1988 Apr;7(4):1139–1145. doi: 10.1002/j.1460-2075.1988.tb02923.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eilers M., Oppliger W., Schatz G. Both ATP and an energized inner membrane are required to import a purified precursor protein into mitochondria. EMBO J. 1987 Apr;6(4):1073–1077. doi: 10.1002/j.1460-2075.1987.tb04860.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eilers M., Schatz G. Binding of a specific ligand inhibits import of a purified precursor protein into mitochondria. Nature. 1986 Jul 17;322(6076):228–232. doi: 10.1038/322228a0. [DOI] [PubMed] [Google Scholar]
  8. Eilers M., Schatz G. Protein unfolding and the energetics of protein translocation across biological membranes. Cell. 1988 Feb 26;52(4):481–483. doi: 10.1016/0092-8674(88)90458-8. [DOI] [PubMed] [Google Scholar]
  9. Ellis J. Proteins as molecular chaperones. 1987 Jul 30-Aug 5Nature. 328(6129):378–379. doi: 10.1038/328378a0. [DOI] [PubMed] [Google Scholar]
  10. Flügge U. I., Hinz G. Energy dependence of protein translocation into chloroplasts. Eur J Biochem. 1986 Nov 3;160(3):563–570. doi: 10.1111/j.1432-1033.1986.tb10075.x. [DOI] [PubMed] [Google Scholar]
  11. Gbelská Y., Subík J., Svoboda A., Goffeau A., Kovác L. Intramitochondrial ATP and cell functions: yeast cells depleted of intramitochondrial ATP lose the ability to grow and multiply. Eur J Biochem. 1983 Feb 1;130(2):281–286. doi: 10.1111/j.1432-1033.1983.tb07148.x. [DOI] [PubMed] [Google Scholar]
  12. Hartl F. U., Pfanner N., Nicholson D. W., Neupert W. Mitochondrial protein import. Biochim Biophys Acta. 1989 Jan 18;988(1):1–45. doi: 10.1016/0304-4157(89)90002-6. [DOI] [PubMed] [Google Scholar]
  13. Hayashi S. I., Lin E. C. Purification and properties of glycerol kinase from Escherichia coli. J Biol Chem. 1967 Mar 10;242(5):1030–1035. [PubMed] [Google Scholar]
  14. Heldt H. W., Klingenberg M., Milovancev M. Differences between the ATP-ADP ratios in the mitochondrial matrix and in the extramitochondrial space. Eur J Biochem. 1972 Nov 7;30(3):434–440. doi: 10.1111/j.1432-1033.1972.tb02115.x. [DOI] [PubMed] [Google Scholar]
  15. Hurt E. C., Pesold-Hurt B., Schatz G. The amino-terminal region of an imported mitochondrial precursor polypeptide can direct cytoplasmic dihydrofolate reductase into the mitochondrial matrix. EMBO J. 1984 Dec 20;3(13):3149–3156. doi: 10.1002/j.1460-2075.1984.tb02272.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hurt E. C., Pesold-Hurt B., Schatz G. The cleavable prepiece of an imported mitochondrial protein is sufficient to direct cytosolic dihydrofolate reductase into the mitochondrial matrix. FEBS Lett. 1984 Dec 10;178(2):306–310. doi: 10.1016/0014-5793(84)80622-5. [DOI] [PubMed] [Google Scholar]
  17. Hwang S., Jascur T., Vestweber D., Pon L., Schatz G. Disrupted yeast mitochondria can import precursor proteins directly through their inner membrane. J Cell Biol. 1989 Aug;109(2):487–493. doi: 10.1083/jcb.109.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Klingenberg M. Molecular aspects of the adenine nucleotide carrier from mitochondria. Arch Biochem Biophys. 1989 Apr;270(1):1–14. doi: 10.1016/0003-9861(89)90001-5. [DOI] [PubMed] [Google Scholar]
  19. Lill R., Cunningham K., Brundage L. A., Ito K., Oliver D., Wickner W. SecA protein hydrolyzes ATP and is an essential component of the protein translocation ATPase of Escherichia coli. EMBO J. 1989 Mar;8(3):961–966. doi: 10.1002/j.1460-2075.1989.tb03458.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nelson N., Schatz G. Energy-dependent processing of cytoplasmically made precursors to mitochondrial proteins. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4365–4369. doi: 10.1073/pnas.76.9.4365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Olsen L. J., Theg S. M., Selman B. R., Keegstra K. ATP is required for the binding of precursor proteins to chloroplasts. J Biol Chem. 1989 Apr 25;264(12):6724–6729. [PubMed] [Google Scholar]
  22. Pain D., Blobel G. Protein import into chloroplasts requires a chloroplast ATPase. Proc Natl Acad Sci U S A. 1987 May;84(10):3288–3292. doi: 10.1073/pnas.84.10.3288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pfanner N., Neupert W. Transport of F1-ATPase subunit beta into mitochondria depends on both a membrane potential and nucleoside triphosphates. FEBS Lett. 1986 Dec 15;209(2):152–156. doi: 10.1016/0014-5793(86)81101-2. [DOI] [PubMed] [Google Scholar]
  24. Pfanner N., Pfaller R., Kleene R., Ito M., Tropschug M., Neupert W. Role of ATP in mitochondrial protein import. Conformational alteration of a precursor protein can substitute for ATP requirement. J Biol Chem. 1988 Mar 25;263(9):4049–4051. [PubMed] [Google Scholar]
  25. Pfanner N., Tropschug M., Neupert W. Mitochondrial protein import: nucleoside triphosphates are involved in conferring import-competence to precursors. Cell. 1987 Jun 19;49(6):815–823. doi: 10.1016/0092-8674(87)90619-2. [DOI] [PubMed] [Google Scholar]
  26. Subík J., Takácsová G., Kovác L. Intramitochondrial ATP and cell functions. I. Growing yeast cells depleted of intramitochondrial ATP are losing mitochondrial genes. Mol Gen Genet. 1978 Oct 25;166(1):103–116. doi: 10.1007/BF00379735. [DOI] [PubMed] [Google Scholar]
  27. Theg S. M., Bauerle C., Olsen L. J., Selman B. R., Keegstra K. Internal ATP is the only energy requirement for the translocation of precursor proteins across chloroplastic membranes. J Biol Chem. 1989 Apr 25;264(12):6730–6736. [PubMed] [Google Scholar]
  28. Verner K., Schatz G. Protein translocation across membranes. Science. 1988 Sep 9;241(4871):1307–1313. doi: 10.1126/science.2842866. [DOI] [PubMed] [Google Scholar]

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