Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

Molecular Biology of the Cell logoLink to Molecular Biology of the Cell
. 1995 Jun;6(6):675–683. doi: 10.1091/mbc.6.6.675

Import of stably folded proteins into peroxisomes.

P A Walton 1, P E Hill 1, S Subramani 1
PMCID: PMC301228  PMID: 7579687

Abstract

By virtue of their synthesis in the cytoplasm, proteins destined for import into peroxisomes are obliged to traverse the single membrane of this organelle. Because the targeting signal for most peroxisomal matrix proteins is a carboxy-terminal tripeptide sequence (SKL or its variants), these proteins must remain import competent until their translation is complete. We sought to determine whether stably folded proteins were substrates for peroxisomal import. Prefolded proteins stabilized with disulfide bonds and chemical cross-linkers were shown to be substrates for peroxisomal import, as were mature folded and disulfide-bonded IgG molecules containing the peroxisomal targeting signal. In addition, colloidal gold particles conjugated to proteins bearing the peroxisomal targeting signal were translocated into the peroxisomal matrix. These results support the concept that proteins may fold in the mammalian cytosol, before their import into the peroxisome, and that protein unfolding is not a prerequisite for peroxisomal import.

Full text

PDF
675

Images in this article

Selected References

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

  1. Bellion E., Goodman J. M. Proton ionophores prevent assembly of a peroxisomal protein. Cell. 1987 Jan 16;48(1):165–173. doi: 10.1016/0092-8674(87)90367-9. [DOI] [PubMed] [Google Scholar]
  2. Brul S., Westerveld A., Strijland A., Wanders R. J., Schram A. W., Heymans H. S., Schutgens R. B., van den Bosch H., Tager J. M. Genetic heterogeneity in the cerebrohepatorenal (Zellweger) syndrome and other inherited disorders with a generalized impairment of peroxisomal functions. A study using complementation analysis. J Clin Invest. 1988 Jun;81(6):1710–1715. doi: 10.1172/JCI113510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Della-Cioppa G., Kishore G. M. Import of a precursor protein into chloroplasts is inhibited by the herbicide glyphosate. EMBO J. 1988 May;7(5):1299–1305. doi: 10.1002/j.1460-2075.1988.tb02944.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dworetzky S. I., Feldherr C. M. Translocation of RNA-coated gold particles through the nuclear pores of oocytes. J Cell Biol. 1988 Mar;106(3):575–584. doi: 10.1083/jcb.106.3.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Feldherr C. M., Kallenbach E., Schultz N. Movement of a karyophilic protein through the nuclear pores of oocytes. J Cell Biol. 1984 Dec;99(6):2216–2222. doi: 10.1083/jcb.99.6.2216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Frydman J., Nimmesgern E., Erdjument-Bromage H., Wall J. S., Tempst P., Hartl F. U. Function in protein folding of TRiC, a cytosolic ring complex containing TCP-1 and structurally related subunits. EMBO J. 1992 Dec;11(13):4767–4778. doi: 10.1002/j.1460-2075.1992.tb05582.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glover J. R., Andrews D. W., Rachubinski R. A. Saccharomyces cerevisiae peroxisomal thiolase is imported as a dimer. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10541–10545. doi: 10.1073/pnas.91.22.10541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hachiya N., Alam R., Sakasegawa Y., Sakaguchi M., Mihara K., Omura T. A mitochondrial import factor purified from rat liver cytosol is an ATP-dependent conformational modulator for precursor proteins. EMBO J. 1993 Apr;12(4):1579–1586. doi: 10.1002/j.1460-2075.1993.tb05802.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hannavy K., Rospert S., Schatz G. Protein import into mitochondria: a paradigm for the translocation of polypeptides across membranes. Curr Opin Cell Biol. 1993 Aug;5(4):694–700. doi: 10.1016/0955-0674(93)90142-d. [DOI] [PubMed] [Google Scholar]
  11. Hwang C., Sinskey A. J., Lodish H. F. Oxidized redox state of glutathione in the endoplasmic reticulum. Science. 1992 Sep 11;257(5076):1496–1502. doi: 10.1126/science.1523409. [DOI] [PubMed] [Google Scholar]
  12. Labarca P., Wolff D., Soto U., Necochea C., Leighton F. Large cation-selective pores from rat liver peroxisomal membranes incorporated to planar lipid bilayers. J Membr Biol. 1986;94(3):285–291. doi: 10.1007/BF01869724. [DOI] [PubMed] [Google Scholar]
  13. Lemmens M., Verheyden K., Van Veldhoven P., Vereecke J., Mannaerts G. P., Carmeliet E. Single-channel analysis of a large conductance channel in peroxisomes from rat liver. Biochim Biophys Acta. 1989 Sep 18;984(3):351–359. doi: 10.1016/0005-2736(89)90302-7. [DOI] [PubMed] [Google Scholar]
  14. McNew J. A., Goodman J. M. An oligomeric protein is imported into peroxisomes in vivo. J Cell Biol. 1994 Dec;127(5):1245–1257. doi: 10.1083/jcb.127.5.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Middelkoop E., Strijland A., Tager J. M. Does aminotriazole inhibit import of catalase into peroxisomes by retarding unfolding? FEBS Lett. 1991 Feb 11;279(1):79–82. doi: 10.1016/0014-5793(91)80255-2. [DOI] [PubMed] [Google Scholar]
  16. Nicolay K., Veenhuis M., Douma A. C., Harder W. A 31P NMR study of the internal pH of yeast peroxisomes. Arch Microbiol. 1987 Feb;147(1):37–41. doi: 10.1007/BF00492902. [DOI] [PubMed] [Google Scholar]
  17. Pfanner N., Neupert W. The mitochondrial protein import apparatus. Annu Rev Biochem. 1990;59:331–353. doi: 10.1146/annurev.bi.59.070190.001555. [DOI] [PubMed] [Google Scholar]
  18. Schleyer M., Neupert W. Transport of proteins into mitochondria: translocational intermediates spanning contact sites between outer and inner membranes. Cell. 1985 Nov;43(1):339–350. doi: 10.1016/0092-8674(85)90039-x. [DOI] [PubMed] [Google Scholar]
  19. Skerjanc I. S., Sheffield W. P., Randall S. K., Silvius J. R., Shore G. C. Import of precursor proteins into mitochondria: site of polypeptide unfolding. J Biol Chem. 1990 Jun 5;265(16):9444–9451. [PubMed] [Google Scholar]
  20. Slot J. W., Geuze H. J. A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol. 1985 Jul;38(1):87–93. [PubMed] [Google Scholar]
  21. Soto U., Pepperkok R., Ansorge W., Just W. W. Import of firefly luciferase into mammalian peroxisomes in vivo requires nucleoside triphosphates. Exp Cell Res. 1993 Mar;205(1):66–75. doi: 10.1006/excr.1993.1059. [DOI] [PubMed] [Google Scholar]
  22. Subramani S. Protein import into peroxisomes and biogenesis of the organelle. Annu Rev Cell Biol. 1993;9:445–478. doi: 10.1146/annurev.cb.09.110193.002305. [DOI] [PubMed] [Google Scholar]
  23. Van Veldhoven P. P., Just W. W., Mannaerts G. P. Permeability of the peroxisomal membrane to cofactors of beta-oxidation. Evidence for the presence of a pore-forming protein. J Biol Chem. 1987 Mar 25;262(9):4310–4318. [PubMed] [Google Scholar]
  24. Vestweber D., Schatz G. A chimeric mitochondrial precursor protein with internal disulfide bridges blocks import of authentic precursors into mitochondria and allows quantitation of import sites. J Cell Biol. 1988 Dec;107(6 Pt 1):2037–2043. doi: 10.1083/jcb.107.6.2037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vestweber D., Schatz G. DNA-protein conjugates can enter mitochondria via the protein import pathway. Nature. 1989 Mar 9;338(6211):170–172. doi: 10.1038/338170a0. [DOI] [PubMed] [Google Scholar]
  26. Vestweber D., Schatz G. Point mutations destabilizing a precursor protein enhance its post-translational import into mitochondria. EMBO J. 1988 Apr;7(4):1147–1151. doi: 10.1002/j.1460-2075.1988.tb02924.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Walton P. A., Gould S. J., Rachubinski R. A., Subramani S., Feramisco J. R. Transport of microinjected alcohol oxidase from Pichia pastoris into vesicles in mammalian cells: involvement of the peroxisomal targeting signal. J Cell Biol. 1992 Aug;118(3):499–508. doi: 10.1083/jcb.118.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Walton P. A., Wendland M., Subramani S., Rachubinski R. A., Welch W. J. Involvement of 70-kD heat-shock proteins in peroxisomal import. J Cell Biol. 1994 Jun;125(5):1037–1046. doi: 10.1083/jcb.125.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wanders R. J., Kos M., Roest B., Meijer A. J., Schrakamp G., Heymans H. S., Tegelaers W. H., van den Bosch H., Schutgens R. B., Tager J. M. Activity of peroxisomal enzymes and intracellular distribution of catalase in Zellweger syndrome. Biochem Biophys Res Commun. 1984 Sep 28;123(3):1054–1061. doi: 10.1016/s0006-291x(84)80240-5. [DOI] [PubMed] [Google Scholar]

Articles from Molecular Biology of the Cell are provided here courtesy of American Society for Cell Biology

RESOURCES