Abstract
A lipoprotein mutant of E. coli K-12 has been characterized. The mutant lipoprotein was found to differ from the wild-type lipoprotein in the following respects: (i) it is present in an appreciable amount in the soluble fraction (275,000 X g supernatant); (ii) it lacks the covalently-linked diglyceride; (iii) it contains an unmodified cysteine which can be carboxymethylated in vitro; (iv) it undergoes dimerization and the dimer can be converted into monomeric form by reduction with 2-mercaptoethanol; (v) both the monomeric form and especially the dimeric form of the mutant lipoprotein migrate more slowly than the corresponding forms of wild-type lipoprotein in sodium dodecyl sulfate/urea polyacrylamide gel electrophoresis; and (vi) the mutant lipoprotein is not assembled into the murein sacculi, and this results in a greatly reduced amount of bound-form lipoprotein in the mutant. These data strongly suggest that the mutation has affected the primary structure of lipoprotein, in such a way that it is not modified normally, leading to the production of a structurally-altered lipoprotein deficient in covalently-linked lipid as well as a defective assembly of the altered lipoprotein into the rigid layer of the cell envelope.
Full text
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Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Braun V., Rehn K. Chemical characterization, spatial distribution and function of a lipoprotein (murein-lipoprotein) of the E. coli cell wall. The specific effect of trypsin on the membrane structure. Eur J Biochem. 1969 Oct;10(3):426–438. doi: 10.1111/j.1432-1033.1969.tb00707.x. [DOI] [PubMed] [Google Scholar]
- 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]
- Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Halegoua S., Hirashima A., Inouye M. Puromycin-resistant biosynthesis of a specific outer-membrane lipoprotein of Escherichia coli. J Bacteriol. 1976 Apr;126(1):183–191. doi: 10.1128/jb.126.1.183-191.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Inouye M., Guthrie J. P. A mutation which changes a membrane protein of E. coli. Proc Natl Acad Sci U S A. 1969 Nov;64(3):957–961. doi: 10.1073/pnas.64.3.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Inoyye S., Takeishi K., Lee N., DeMartini M., Hirashima A., Inouye M. Lipoprotein from the outer membrane of Escherichia coli: purification, paracrystallization, and some properties of its free form. J Bacteriol. 1976 Jul;127(1):555–563. doi: 10.1128/jb.127.1.555-563.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin J. J., Wu H. C. Biosynthesis and assembly of envelope lipoprotein in a glycerol-requiring mutant of Salmonella typhimurium. J Bacteriol. 1976 Mar;125(3):892–904. doi: 10.1128/jb.125.3.892-904.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moldow C., Robertson J., Rothfield L. Purification of bacterial membrane proteins. The use of guanidinium thiocyanate and urea. J Membr Biol. 1972;10(2):137–152. doi: 10.1007/BF01867850. [DOI] [PubMed] [Google Scholar]
- Ozols J. Amino acid sequence of rabbit liver microsomal cytochrome b5. J Biol Chem. 1970 Oct 10;245(19):4863–4874. [PubMed] [Google Scholar]
- Suzuki H., Nishimura Y., Iketani H., Campisi J., Hirashima A. Novel mutation that causes a structural change in a lipoprotein in the outer membrane of Escherichia coli. J Bacteriol. 1976 Sep;127(3):1494–1501. doi: 10.1128/jb.127.3.1494-1501.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
- Wu H. C., Lin J. J. Escherichia coli mutants altered in murein lipoprotein. J Bacteriol. 1976 Apr;126(1):147–156. doi: 10.1128/jb.126.1.147-156.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]