Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Oct;71(10):4149–4153. doi: 10.1073/pnas.71.10.4149

Cell-free Synthesis of a Specific Lipoprotein of the Escherichia coli Outer Membrane Directed by Purified Messenger RNA

Akikazu Hirashima 1, Sansan Wang 1, Masayori Inouye 1
PMCID: PMC434347  PMID: 4610570

Abstract

A specific lipoprotein of the E. coli outer membrane has been synthesized in a cell-free system directed by purified messenger RNA. The mRNA for the lipoprotein was purified as 7S RNA about 250-fold from exponentially growing cells. Protein synthesis of the cell-free system was totally dependent upon the addition of the purified mRNA. The product of the cell-free system was identified as the specific lipoprotein by immunoprecipitation and by peptide mapping.

Keywords: immunoassay, BrCN cleavage, protein identification

Full text

PDF
4149

Images in this article

4151Image
on p.4151
4152Image
on p.4152

Selected References

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

  1. Adams J. M. On the release of the formyl group from nascent protein. J Mol Biol. 1968 May 14;33(3):571–589. doi: 10.1016/0022-2836(68)90307-0. [DOI] [PubMed] [Google Scholar]
  2. Bosch V., Braun V. Distribution of murein-lipoprotein between the cytoplasmic and outer membrane of Escherichia coli. FEBS Lett. 1973 Aug 15;34(2):307–310. doi: 10.1016/0014-5793(73)80818-x. [DOI] [PubMed] [Google Scholar]
  3. Braun V., Bosch V. Repetitive sequences in the murein-lipoprotein of the cell wall of Escherichia coli. Proc Natl Acad Sci U S A. 1972 Apr;69(4):970–974. doi: 10.1073/pnas.69.4.970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Braun V., Bosch V. Sequence of the murein-lipoprotein and the attachment site of the lipid. Eur J Biochem. 1972 Jun 23;28(1):51–69. doi: 10.1111/j.1432-1033.1972.tb01883.x. [DOI] [PubMed] [Google Scholar]
  5. Braun V., Sieglin U. The covalent murein-lipoprotein structure of the Escherichia coli cell wall. The attachment site of the lipoprotein on the murein. Eur J Biochem. 1970 Apr;13(2):336–346. doi: 10.1111/j.1432-1033.1970.tb00936.x. [DOI] [PubMed] [Google Scholar]
  6. Braun V., Wolff H. The murein-lipoprotein linkage in the cell wall of Escherichia coli. Eur J Biochem. 1970 Jun;14(2):387–391. doi: 10.1111/j.1432-1033.1970.tb00301.x. [DOI] [PubMed] [Google Scholar]
  7. Bronson M. J., Squires C., Yanofsky C. Nucleotide sequences from tryptophan messenger RNA of Escherichia coli: the sequence corresponding to the amino-terminal region of the first polypeptide specified by the operon. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2335–2339. doi: 10.1073/pnas.70.8.2335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Capecchi M. R. Initiation of E. coli proteins. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1517–1524. doi: 10.1073/pnas.55.6.1517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dudock B. S., Katz G., Taylor E. K., Holley R. W. Primary structure of wheat germ phenylalanine transfer RNA. Proc Natl Acad Sci U S A. 1969 Mar;62(3):941–945. doi: 10.1073/pnas.62.3.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Forchhammer J., Lindahl L. Growth rate of polypeptide chains as a function of the cell growth rate in a mutant of Escherichia coli 15. J Mol Biol. 1971 Feb 14;55(3):563–568. doi: 10.1016/0022-2836(71)90337-8. [DOI] [PubMed] [Google Scholar]
  11. Gausing K. Efficiency of protein and messenger RNA synthesis in bacteriophage T4-infected cells of Escherichia coli. J Mol Biol. 1972 Nov 28;71(3):529–545. doi: 10.1016/s0022-2836(72)80021-4. [DOI] [PubMed] [Google Scholar]
  12. Hantke K., Braun V. Covalent binding of lipid to protein. Diglyceride and amide-linked fatty acid at the N-terminal end of the murein-lipoprotein of the Escherichia coli outer membrane. Eur J Biochem. 1973 Apr;34(2):284–296. doi: 10.1111/j.1432-1033.1973.tb02757.x. [DOI] [PubMed] [Google Scholar]
  13. Hirashima A., Childs G., Inouye M. Differential inhibitory effects of antibiotics on the biosynthesis of envelope proteins of Escherichia coli. J Mol Biol. 1973 Sep 15;79(2):373–389. doi: 10.1016/0022-2836(73)90012-0. [DOI] [PubMed] [Google Scholar]
  14. Hirashima A., Inouye M. Specific biosynthesis of an envelope protein of Escherichia coli. Nature. 1973 Apr 6;242(5397):405–407. doi: 10.1038/242405a0. [DOI] [PubMed] [Google Scholar]
  15. Hirashima A., Wu H. C., Venkateswaran P. S., Inouye M. Two forms of a structural lipoprotein in the envelope of Escherichia coli. Further characterization of the free form. J Biol Chem. 1973 Aug 25;248(16):5654–5659. [PubMed] [Google Scholar]
  16. Inouye M. A three-dimensional molecular assembly model of a lipoprotein from the Escherichia coli outer membrane. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2396–2400. doi: 10.1073/pnas.71.6.2396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Inouye M. Internal standards for molecular weight determinations of proteins by polyacrylamide gel electrophoresis. Applications to envelope proteins of Escherichia coli. J Biol Chem. 1971 Aug 10;246(15):4834–4838. [PubMed] [Google Scholar]
  18. Inouye M., Pardee A. B. Changes of membrane proteins and their relation to deoxyribonucleic acid synthesis and cell division of Escherichia coli. J Biol Chem. 1970 Nov 10;245(21):5813–5819. [PubMed] [Google Scholar]
  19. Inouye M., Shaw J., Shen C. The assembly of a structural lipoprotein in the envelope of Escherichia coli. J Biol Chem. 1972 Dec 25;247(24):8154–8159. [PubMed] [Google Scholar]
  20. Lacroute F., Stent G. S. Peptide chain growth of -galactosidase in Escherichia coli. J Mol Biol. 1968 Jul 14;35(1):165–173. doi: 10.1016/s0022-2836(68)80044-0. [DOI] [PubMed] [Google Scholar]
  21. Lee N., Inouye M. Outer membrane proteins of Escherichia coli: biosynthesis and assembly. FEBS Lett. 1974 Feb 15;39(2):167–170. doi: 10.1016/0014-5793(74)80043-8. [DOI] [PubMed] [Google Scholar]
  22. Maizels N. M. The nucleotide sequence of the lactose messenger ribonucleic acid transcribed from the UV5 promoter mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3585–3589. doi: 10.1073/pnas.70.12.3585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. NIRENBERG M. W., MATTHAEI J. H. The dependence of cell-free protein synthesis in E. coli upon naturally occurring or synthetic polyribonucleotides. Proc Natl Acad Sci U S A. 1961 Oct 15;47:1588–1602. doi: 10.1073/pnas.47.10.1588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Peacock A. C., Dingman C. W. Resolution of multiple ribonucleic acid species by polyacrylamide gel electrophoresis. Biochemistry. 1967 Jun;6(6):1818–1827. doi: 10.1021/bi00858a033. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES