Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1989 Dec 1;8(12):3917–3921. doi: 10.1002/j.1460-2075.1989.tb08572.x

The reaction specificities of the thylakoidal processing peptidase and Escherichia coli leader peptidase are identical.

C Halpin 1, P D Elderfield 1, H E James 1, R Zimmermann 1, B Dunbar 1, C Robinson 1
PMCID: PMC402083  PMID: 2684650

Abstract

Proteins which are transported across the bacterial plasma membrane, endoplasmic reticulum and thylakoid membrane are usually synthesized as larger precursors containing amino-terminal targeting signals. Removal of the signals is carried out by specific, membrane-bound processing peptidases. In this report we show that the reaction specificities of these three peptidases are essentially identical. Precursors of two higher plant thylakoid lumen proteins are efficiently processed by purified Escherichia coli leader peptidase. Processing of one precursor, that of the 23 kd photosystem II protein, by both the thylakoidal and E. coli enzymes generates the correct mature amino terminus. Similarly, leader (signal) peptides of both eukaryotic and prokaryotic origin are cleaved by partially purified thylakoidal processing peptidase. No evidence of incorrect processing was obtained. Both leader peptidase and thylakoidal peptidase are inhibited by a synthetic leader peptide.

Full text

PDF

Images in this article

3918Image
on p.3918
3918Image
on p.3918
3919Image
on p.3919
3919Image
on p.3919

Selected References

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

  1. Anderson C. W., Straus J. W., Dudock B. S. Preparation of a cell-free protein-synthesizing system from wheat germ. Methods Enzymol. 1983;101:635–644. doi: 10.1016/0076-6879(83)01044-7. [DOI] [PubMed] [Google Scholar]
  2. Austen B. M., Hermon-Taylor J., Kaderbhai M. A., Ridd D. H. Design and synthesis of a consensus signal sequence that inhibits protein translocation into rough microsomal vesicles. Biochem J. 1984 Nov 15;224(1):317–325. doi: 10.1042/bj2240317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker R. K., Lively M. O. Purification and characterization of hen oviduct microsomal signal peptidase. Biochemistry. 1987 Dec 29;26(26):8561–8567. doi: 10.1021/bi00400a010. [DOI] [PubMed] [Google Scholar]
  4. Böhni P. C., Deshaies R. J., Schekman R. W. SEC11 is required for signal peptide processing and yeast cell growth. J Cell Biol. 1988 Apr;106(4):1035–1042. doi: 10.1083/jcb.106.4.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crooke E., Guthrie B., Lecker S., Lill R., Wickner W. ProOmpA is stabilized for membrane translocation by either purified E. coli trigger factor or canine signal recognition particle. Cell. 1988 Sep 23;54(7):1003–1011. doi: 10.1016/0092-8674(88)90115-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans E. A., Gilmore R., Blobel G. Purification of microsomal signal peptidase as a complex. Proc Natl Acad Sci U S A. 1986 Feb;83(3):581–585. doi: 10.1073/pnas.83.3.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Hawlitschek G., Schneider H., Schmidt B., Tropschug M., Hartl F. U., Neupert W. Mitochondrial protein import: identification of processing peptidase and of PEP, a processing enhancing protein. Cell. 1988 Jun 3;53(5):795–806. doi: 10.1016/0092-8674(88)90096-7. [DOI] [PubMed] [Google Scholar]
  9. Jackson R. C., Blobel G. Post-translational cleavage of presecretory proteins with an extract of rough microsomes from dog pancreas containing signal peptidase activity. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5598–5602. doi: 10.1073/pnas.74.12.5598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kirwin P. M., Elderfield P. D., Robinson C. Transport of proteins into chloroplasts. Partial purification of a thylakoidal processing peptidase involved in plastocyanin biogenesis. J Biol Chem. 1987 Dec 5;262(34):16386–16390. [PubMed] [Google Scholar]
  11. Kirwin P. M., Elderfield P. D., Williams R. S., Robinson C. Transport of proteins into chloroplasts. Organization, orientation, and lateral distribution of the plastocyanin processing peptidase in the thylakoid network. J Biol Chem. 1988 Dec 5;263(34):18128–18132. [PubMed] [Google Scholar]
  12. Kirwin P. M., Meadows J. W., Shackleton J. B., Musgrove J. E., Elderfield P. D., Mould R., Hay N. A., Robinson C. ATP-dependent import of a lumenal protein by isolated thylakoid vesicles. EMBO J. 1989 Aug;8(8):2251–2255. doi: 10.1002/j.1460-2075.1989.tb08349.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kronenberg H. M., Fennick B. J., Vasicek T. J. Transport and cleavage of bacterial pre-beta-lactamase by mammalian microsomes. J Cell Biol. 1983 Apr;96(4):1117–1119. doi: 10.1083/jcb.96.4.1117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kurjan J., Herskowitz I. Structure of a yeast pheromone gene (MF alpha): a putative alpha-factor precursor contains four tandem copies of mature alpha-factor. Cell. 1982 Oct;30(3):933–943. doi: 10.1016/0092-8674(82)90298-7. [DOI] [PubMed] [Google Scholar]
  15. Kuwabara T., Reddy K. J., Sherman L. A. Nucleotide sequence of the gene from the cyanobacterium Anacystis nidulans R2 encoding the Mn-stabilizing protein involved in photosystem II water oxidation. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8230–8234. doi: 10.1073/pnas.84.23.8230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lei S. P., Lin H. C., Wang S. S., Callaway J., Wilcox G. Characterization of the Erwinia carotovora pelB gene and its product pectate lyase. J Bacteriol. 1987 Sep;169(9):4379–4383. doi: 10.1128/jb.169.9.4379-4383.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Müller M., Ibrahimi I., Chang C. N., Walter P., Blobel G. A bacterial secretory protein requires signal recognition particle for translocation across mammalian endoplasmic reticulum. J Biol Chem. 1982 Oct 25;257(20):11860–11863. [PubMed] [Google Scholar]
  19. Robinson C., Ellis R. J. Transport of proteins into chloroplasts. Partial purification of a chloroplast protease involved in the processing of important precursor polypeptides. Eur J Biochem. 1984 Jul 16;142(2):337–342. doi: 10.1111/j.1432-1033.1984.tb08291.x. [DOI] [PubMed] [Google Scholar]
  20. Shelness G. S., Kanwar Y. S., Blobel G. cDNA-derived primary structure of the glycoprotein component of canine microsomal signal peptidase complex. J Biol Chem. 1988 Nov 15;263(32):17063–17070. [PubMed] [Google Scholar]
  21. Smeekens S., Bauerle C., Hageman J., Keegstra K., Weisbeek P. The role of the transit peptide in the routing of precursors toward different chloroplast compartments. Cell. 1986 Aug 1;46(3):365–375. doi: 10.1016/0092-8674(86)90657-4. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Wallace T. P., Stewart A. C., Pappin D., Howe C. J. Gene sequence for the 9 kDa component of Photosystem II from the cyanobacterium Phormidium laminosum indicates similarities between cyanobacterial and other leader sequences. Mol Gen Genet. 1989 Apr;216(2-3):334–339. doi: 10.1007/BF00334373. [DOI] [PubMed] [Google Scholar]
  24. Watts C., Wickner W., Zimmermann R. M13 procoat and a pre-immunoglobulin share processing specificity but use different membrane receptor mechanisms. Proc Natl Acad Sci U S A. 1983 May;80(10):2809–2813. doi: 10.1073/pnas.80.10.2809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wickner W., Moore K., Dibb N., Geissert D., Rice M. Inhibition of purified Escherichia coli leader peptidase by the leader (signal) peptide of bacteriophage M13 procoat. J Bacteriol. 1987 Aug;169(8):3821–3822. doi: 10.1128/jb.169.8.3821-3822.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wolfe P. B., Silver P., Wickner W. The isolation of homogeneous leader peptidase from a strain of Escherichia coli which overproduces the enzyme. J Biol Chem. 1982 Jul 10;257(13):7898–7902. [PubMed] [Google Scholar]
  27. Wolfe P. B., Wickner W., Goodman J. M. Sequence of the leader peptidase gene of Escherichia coli and the orientation of leader peptidase in the bacterial envelope. J Biol Chem. 1983 Oct 10;258(19):12073–12080. [PubMed] [Google Scholar]
  28. Zimmermann R., Mollay C. Import of honeybee prepromelittin into the endoplasmic reticulum. Requirements for membrane insertion, processing, and sequestration. J Biol Chem. 1986 Sep 25;261(27):12889–12895. [PubMed] [Google Scholar]
  29. Zimmermann R., Watts C., Wickner W. The biosynthesis of membrane-bound M13 coat protein. Energetics and assembly intermediates. J Biol Chem. 1982 Jun 10;257(11):6529–6536. [PubMed] [Google Scholar]
  30. Zwizinski C., Wickner W. Purification and characterization of leader (signal) peptidase from Escherichia coli. J Biol Chem. 1980 Aug 25;255(16):7973–7977. [PubMed] [Google Scholar]
  31. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]
  32. von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. doi: 10.1016/0022-2836(85)90046-4. [DOI] [PubMed] [Google Scholar]
  33. von Heijne G., Steppuhn J., Herrmann R. G. Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem. 1989 Apr 1;180(3):535–545. doi: 10.1111/j.1432-1033.1989.tb14679.x. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES