Abstract
Lysozyme digestion and sonication of sodium dodecyl sulfate (SDS)-purified Klebsiella aerogenes murein sacculi resulted in the quantitative release of both subunits of nitrate reductase, as well as a number of other cytoplasmic membrane polypeptides (5.2%, by weight, of the total membrane proteins). Similar results were obtained after lysozyme digestion of SDS-prepared peptidoglycan fragments, which excluded the phenomenon of simple trapping of the polypeptides by the surrounding peptidoglycan matrix. About 28% of membrane-bound nitrate reductase appears to be tightly associated with the peptidoglycan. Additional evidence for this association was demonstrated by positive immunogold labeling of SDS-murein sacculi and thin sections of plasmolyzed bacteria. Qualitative amino acid analysis of trypsin-treated sacculi, a tryptic product of holo-nitrate reductase, and amino- and carboxypeptidase digests of both nitrate reductase subunits indicated the possible existence of a terminal anchoring peptide containing the following amino acids: (Gly)n, Trp, Ser, Pro, Ile, Leu, Phe, Cys, Tyr, Asp, and Lys.
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- 't Riet J van, Stouthamer A. H., Planta R. J. Regulation of nitrate assimilation and nitrate respiration in Aerobacter aerogenes. J Bacteriol. 1968 Nov;96(5):1455–1464. doi: 10.1128/jb.96.5.1455-1464.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bayer M. E. Areas of adhesion between wall and membrane of Escherichia coli. J Gen Microbiol. 1968 Oct;53(3):395–404. doi: 10.1099/00221287-53-3-395. [DOI] [PubMed] [Google Scholar]
- Bayer M. H., Costello G. P., Bayer M. E. Isolation and partial characterization of membrane vesicles carrying markers of the membrane adhesion sites. J Bacteriol. 1982 Feb;149(2):758–767. doi: 10.1128/jb.149.2.758-767.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birdsell D. C., Cota-Robles E. H. Production and ultrastructure of lysozyme and ethylenediaminetetraacetate-lysozyme spheroplasts of Escherichia coli. J Bacteriol. 1967 Jan;93(1):427–437. doi: 10.1128/jb.93.1.427-437.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bosma H. J., Wever R., van 't Riet J. Electron paramagnetic resonance studies on membrane-bound respiratory nitrate reductase of Klebsiella aerogenes. FEBS Lett. 1978 Jun 1;90(1):107–111. doi: 10.1016/0014-5793(78)80308-1. [DOI] [PubMed] [Google Scholar]
- Braun V. Covalent lipoprotein from the outer membrane of Escherichia coli. Biochim Biophys Acta. 1975 Oct 31;415(3):335–377. doi: 10.1016/0304-4157(75)90013-1. [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]
- Chang J. Y., Creaser E. H. Separation of 4-N,N-dimethylaminoazobenzene-4'-sulfonyl amino acids on polyamide sheets. J Chromatogr. 1976 Jan 7;116(1):215–217. doi: 10.1016/s0021-9673(00)83724-3. [DOI] [PubMed] [Google Scholar]
- Forsberg C. W., Costerton J. W., Macleod R. A. Separation and localization of cell wall layers of a gram-negative bacterium. J Bacteriol. 1970 Dec;104(3):1338–1353. doi: 10.1128/jb.104.3.1338-1353.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graham A., Boxer D. H. The organization of formate dehydrogenase in the cytoplasmic membrane of Escherichia coli. Biochem J. 1981 Jun 1;195(3):627–637. doi: 10.1042/bj1950627. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hjelmeland L. M. A nondenaturing zwitterionic detergent for membrane biochemistry: design and synthesis. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6368–6370. doi: 10.1073/pnas.77.11.6368. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hobot J. A., Carlemalm E., Villiger W., Kellenberger E. Periplasmic gel: new concept resulting from the reinvestigation of bacterial cell envelope ultrastructure by new methods. J Bacteriol. 1984 Oct;160(1):143–152. doi: 10.1128/jb.160.1.143-152.1984. [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]
- Kalb V. F., Jr, Bernlohr R. W. A new spectrophotometric assay for protein in cell extracts. Anal Biochem. 1977 Oct;82(2):362–371. doi: 10.1016/0003-2697(77)90173-7. [DOI] [PubMed] [Google Scholar]
- LEUTGEB W., WEIDEL W. UBER EIN IN COLI-ZELLWANDPRAEPARATEN ZURUECKGEHALTENES GLYKOGEN. Z Naturforsch B. 1963 Dec;18:1060–1062. [PubMed] [Google Scholar]
- Lin J. K., Chang J. Y. Chromophoric labeling of amino acids with 4-dimethylaminoazobenzene-4'-sulfonyl chloride. Anal Chem. 1975 Aug;47(9):1634–1638. doi: 10.1021/ac60359a007. [DOI] [PubMed] [Google Scholar]
- Lugtenberg B., Van Alphen L. Molecular architecture and functioning of the outer membrane of Escherichia coli and other gram-negative bacteria. Biochim Biophys Acta. 1983 Mar 21;737(1):51–115. doi: 10.1016/0304-4157(83)90014-x. [DOI] [PubMed] [Google Scholar]
- Miura T., Mizushima S. Separation and properties of outer and cytoplasmic membranes in Escherichia coli. Biochim Biophys Acta. 1969;193(2):268–276. doi: 10.1016/0005-2736(69)90188-6. [DOI] [PubMed] [Google Scholar]
- Miura T., Mizushima S. Separation by density gradient centrifugation of two types of membranes from spheroplast membrane of Escherichia coli K12. Biochim Biophys Acta. 1968 Jan 3;150(1):159–161. doi: 10.1016/0005-2736(68)90020-5. [DOI] [PubMed] [Google Scholar]
- Nakayama T., Munoz L., Doi R. H. A procedure to remove protease activities from Bacillus subtilis sporulating cells and their crude extracts. Anal Biochem. 1977 Mar;78(1):165–170. doi: 10.1016/0003-2697(77)90020-3. [DOI] [PubMed] [Google Scholar]
- Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
- Petit J. F., Munoz E., Ghuysen J. M. Peptide cross-links in bacterial cell wall peptidoglycans studied with specific endopeptidases from Streptomyces albus G. Biochemistry. 1966 Aug;5(8):2764–2776. doi: 10.1021/bi00872a037. [DOI] [PubMed] [Google Scholar]
- Reusch V. M., Jr Isolation and analysis of sacculi from Streptococcus sanguis. J Bacteriol. 1982 Sep;151(3):1543–1552. doi: 10.1128/jb.151.3.1543-1552.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reynolds J. A., Tanford C. The gross conformation of protein-sodium dodecyl sulfate complexes. J Biol Chem. 1970 Oct 10;245(19):5161–5165. [PubMed] [Google Scholar]
- Riet J van't The participation of cytochromes in the process of nitrate respiration in klesbsiella (Aerobacter) aerogenes. Biochim Biophys Acta. 1973 Jan 18;292(1):237–245. doi: 10.1016/0005-2728(73)90268-5. [DOI] [PubMed] [Google Scholar]
- Rodríguez-Tébar A., Barbas J. A., Vázquez D. Location of some proteins involved in peptidoglycan synthesis and cell division in the inner and outer membranes of Escherichia coli. J Bacteriol. 1985 Jan;161(1):243–248. doi: 10.1128/jb.161.1.243-248.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenbusch J. P. Characterization of the major envelope protein from Escherichia coli. Regular arrangement on the peptidoglycan and unusual dodecyl sulfate binding. J Biol Chem. 1974 Dec 25;249(24):8019–8029. [PubMed] [Google Scholar]
- Slot J. W., Geuze H. J. Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol. 1981 Aug;90(2):533–536. doi: 10.1083/jcb.90.2.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vaessen R. T., Kreike J., Groot G. S. Protein transfer to nitrocellulose filters. A simple method for quantitation of single proteins in complex mixtures. FEBS Lett. 1981 Feb 23;124(2):193–196. doi: 10.1016/0014-5793(81)80134-2. [DOI] [PubMed] [Google Scholar]
- Van 't Riet J., Planta R. J. Purification, structure and properties of the respiratory nitrate reductase of Klebsiella aerogenes. Biochim Biophys Acta. 1975 Jan 30;379(1):81–94. doi: 10.1016/0005-2795(75)90010-0. [DOI] [PubMed] [Google Scholar]
- Verwer R. W., Beachey E. H., Keck W., Stoub A. M., Poldermans J. E. Oriented fragmentation of Escherichia coli sacculi by sonication. J Bacteriol. 1980 Jan;141(1):327–332. doi: 10.1128/jb.141.1.327-332.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wensink J., Witholt B. Identification of different forms of the murein-bound lipoprotein found in isolated outer membranes of Escherichia coli. Eur J Biochem. 1981 Jan;113(2):349–357. doi: 10.1111/j.1432-1033.1981.tb05073.x. [DOI] [PubMed] [Google Scholar]
- Wientjes F. B., Kolk A. H., Nanninga N., Van T'Riet J. Respiratory nitrate reductase: its localization in the cytoplasmic membrane of Klebsiella aerogenes and Bacillus licheniformis. Eur J Biochem. 1979 Mar 15;95(1):61–67. doi: 10.1111/j.1432-1033.1979.tb12939.x. [DOI] [PubMed] [Google Scholar]
- Witholt B., Boekhout M., Brock M., Kingma J., Heerikhuizen H. V., Leij L. D. An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal Biochem. 1976 Jul;74(1):160–170. doi: 10.1016/0003-2697(76)90320-1. [DOI] [PubMed] [Google Scholar]
- Woldringh C. L., de Jong M. A., van den Berg W., Koppes L. Morphological analysis of the division cycle of two Escherichia coli substrains during slow growth. J Bacteriol. 1977 Jul;131(1):270–279. doi: 10.1128/jb.131.1.270-279.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Heij H. T., Jochemsen A. G., Willemsen P. T., Groot G. S. Protein synthesis during chloroplast development in Spirodela oligorhiza. Coordinated synthesis of chloroplast-encoded and nuclear-encoded subunits of ATPase and ribulose-1,5-bisphosphate carboxylase. Eur J Biochem. 1984 Jan 2;138(1):161–168. doi: 10.1111/j.1432-1033.1984.tb07895.x. [DOI] [PubMed] [Google Scholar]





