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. 1986 Mar;165(3):787–795. doi: 10.1128/jb.165.3.787-795.1986

Lateral diffusion of proteins in the periplasm of Escherichia coli.

J M Brass, C F Higgins, M Foley, P A Rugman, J Birmingham, P B Garland
PMCID: PMC214497  PMID: 3005237

Abstract

We have introduced biologically active, fluorescently labeled maltose-binding protein into the periplasmic space of Escherichia coli and measured its lateral diffusion coefficient by the fluorescence photobleaching recovery method. Diffusion of this protein in the periplasm was found to be surprisingly low (lateral diffusion coefficient, 0.9 X 10(-10) cm2 s-1), about 1,000-fold lower than would be expected for diffusion in aqueous medium and almost 100-fold lower than for an equivalent-size protein in the cytoplasm. Galactose-binding protein, myoglobin, and cytochrome c were also introduced into the periplasm and had diffusion coefficients identical to that determined for the maltose-binding protein. For all proteins nearly 100% recovery of fluorescence was obtained after photobleaching, indicating that the periplasm is a single contiguous compartment surrounding the cell. These data have considerable implications for periplasmic structure and for the role of periplasmic proteins in transport and chemotaxis.

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Selected References

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  1. Ames G. F., Spurich E. N. Protein-protein interaction in transport: periplasmic histidine-binding protein J interacts with P protein. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1877–1881. doi: 10.1073/pnas.73.6.1877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Axelrod D., Koppel D. E., Schlessinger J., Elson E., Webb W. W. Mobility measurement by analysis of fluorescence photobleaching recovery kinetics. Biophys J. 1976 Sep;16(9):1055–1069. doi: 10.1016/S0006-3495(76)85755-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bavoil P., Nikaido H. Physical interaction between the phage lambda receptor protein and the carrier-immobilized maltose-binding protein of Escherichia coli. J Biol Chem. 1981 Nov 25;256(22):11385–11388. [PubMed] [Google Scholar]
  4. Boos W., Gordon A. S., Hall R. E., Price H. D. Transport properties of the galactose-binding protein of Escherichia coli. Substrate-induced conformational change. J Biol Chem. 1972 Feb 10;247(3):917–924. [PubMed] [Google Scholar]
  5. Boos W. Structurally defective galactose-binding protein isolated from a mutant negative in the -methylgalactoside transport system of Escherichia coli. J Biol Chem. 1972 Sep 10;247(17):5414–5424. [PubMed] [Google Scholar]
  6. Brass J. M., Boos W., Hengge R. Reconstitution of maltose transport in malB mutants of Escherichia coli through calcium-induced disruptions of the outer membrane. J Bacteriol. 1981 Apr;146(1):10–17. doi: 10.1128/jb.146.1.10-17.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brass J. M., Ehmann U., Bukau B. Reconstitution of maltose transport in Escherichia coli: conditions affecting import of maltose-binding protein into the periplasm of calcium-treated cells. J Bacteriol. 1983 Jul;155(1):97–106. doi: 10.1128/jb.155.1.97-106.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brass J. M., Manson M. D., Larson T. J. Transposon Tn10-dependent expression of the lamB gene in Escherichia coli. J Bacteriol. 1984 Jul;159(1):93–99. doi: 10.1128/jb.159.1.93-99.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Brass J. M., Manson M. D. Reconstitution of maltose chemotaxis in Escherichia coli by addition of maltose-binding protein to calcium-treated cells of maltose regulon mutants. J Bacteriol. 1984 Mar;157(3):881–890. doi: 10.1128/jb.157.3.881-890.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Braun V., Hantke K. Biochemistry of bacterial cell envelopes. Annu Rev Biochem. 1974;43(0):89–121. doi: 10.1146/annurev.bi.43.070174.000513. [DOI] [PubMed] [Google Scholar]
  11. Bukau B., Brass J. M., Boos W. Ca2+-induced permeabilization of the Escherichia coli outer membrane: comparison of transformation and reconstitution of binding-protein-dependent transport. J Bacteriol. 1985 Jul;163(1):61–68. doi: 10.1128/jb.163.1.61-68.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cherry R. J. Rotational and lateral diffusion of membrane proteins. Biochim Biophys Acta. 1979 Dec 20;559(4):289–327. doi: 10.1016/0304-4157(79)90009-1. [DOI] [PubMed] [Google Scholar]
  13. Davison M. T., Garland P. B. Immunochemical demonstration of zonal growth of the cell envelope of Escherichia coli. Eur J Biochem. 1983 Feb 15;130(3):589–597. doi: 10.1111/j.1432-1033.1983.tb07190.x. [DOI] [PubMed] [Google Scholar]
  14. Dietzel I., Kolb V., Boos W. Pole cap formation in Escherichia coli following induction of the maltose-binding protein. Arch Microbiol. 1978 Aug 1;118(2):207–218. doi: 10.1007/BF00415731. [DOI] [PubMed] [Google Scholar]
  15. Ferenci T., Klotz U. Affinity chromatographic isolation of the periplasmic maltose binding protein of Escherichia coli. FEBS Lett. 1978 Oct 15;94(2):213–217. doi: 10.1016/0014-5793(78)80940-5. [DOI] [PubMed] [Google Scholar]
  16. Garland P. Fluorescence photobleaching recovery: control of laser intensities with an acousto-optic modulator. Biophys J. 1981 Mar;33(3):481–482. doi: 10.1016/S0006-3495(81)84910-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hazelbauer G. L., Adler J. Role of the galactose binding protein in chemotaxis of Escherichia coli toward galactose. Nat New Biol. 1971 Mar 24;230(12):101–104. doi: 10.1038/newbio230101a0. [DOI] [PubMed] [Google Scholar]
  18. Hazelbauer G. L. Maltose chemoreceptor of Escherichia coli. J Bacteriol. 1975 Apr;122(1):206–214. doi: 10.1128/jb.122.1.206-214.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hengge R., Boos W. Maltose and lactose transport in Escherichia coli. Examples of two different types of concentrative transport systems. Biochim Biophys Acta. 1983 Aug 11;737(3-4):443–478. doi: 10.1016/0304-4157(83)90009-6. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Hochman J. H., Schindler M., Lee J. G., Ferguson-Miller S. Lateral mobility of cytochrome c on intact mitochondrial membranes as determined by fluorescence redistribution after photobleaching. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6866–6870. doi: 10.1073/pnas.79.22.6866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jacobson K., Wojcieszyn J. The translational mobility of substances within the cytoplasmic matrix. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6747–6751. doi: 10.1073/pnas.81.21.6747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Koiwai O., Hayashi H. Studies on bacterial chemotaxis. IV. Interaction of maltose receptor with a membrane-bound chemosensing component. J Biochem. 1979 Jul;86(1):27–34. [PubMed] [Google Scholar]
  24. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Macalister T. J., Macdonald B., Rothfield L. I. The periseptal annulus: An organelle associated with cell division in Gram-negative bacteria. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1372–1376. doi: 10.1073/pnas.80.5.1372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McCloskey M., Poo M. M. Protein diffusion in cell membranes: some biological implications. Int Rev Cytol. 1984;87:19–81. doi: 10.1016/s0074-7696(08)62439-0. [DOI] [PubMed] [Google Scholar]
  29. Mirelman D., Yashouv-Gan Y., Schwarz U. Regulation of murein biosynthesis and septum formation in filamentous cells of Escherichia coli PAT 84. J Bacteriol. 1977 Mar;129(3):1593–1600. doi: 10.1128/jb.129.3.1593-1600.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mitchell P. Translocations through natural membranes. Adv Enzymol Relat Areas Mol Biol. 1967;29:33–87. doi: 10.1002/9780470122747.ch2. [DOI] [PubMed] [Google Scholar]
  31. Mowbray S. L., Petsko G. A. The x-ray structure of the periplasmic galactose binding protein from Salmonella typhimurium at 3.0-A resolution. J Biol Chem. 1983 Jul 10;258(13):7991–7997. doi: 10.2210/pdb1gbp/pdb. [DOI] [PubMed] [Google Scholar]
  32. Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
  33. Nossal N. G., Heppel L. A. The release of enzymes by osmotic shock from Escherichia coli in exponential phase. J Biol Chem. 1966 Jul 10;241(13):3055–3062. [PubMed] [Google Scholar]
  34. Nozaki Y., Imada A., Yoneda M. SCE-963, a new potent cephalosporin with high affinity for penicillin-binding proteins 1 and 3 of Escherichia coli. Antimicrob Agents Chemother. 1979 Jan;15(1):20–27. doi: 10.1128/aac.15.1.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Peters R. Translational diffusion in the plasma membrane of single cells as studied by fluorescence microphotolysis. Cell Biol Int Rep. 1981 Aug;5(8):733–760. doi: 10.1016/0309-1651(81)90231-9. [DOI] [PubMed] [Google Scholar]
  36. Quiocho F. A., Vyas N. K. Novel stereospecificity of the L-arabinose-binding protein. Nature. 1984 Aug 2;310(5976):381–386. doi: 10.1038/310381a0. [DOI] [PubMed] [Google Scholar]
  37. Raj T., Flygare W. H. Diffusion studies of bovine serum albumin by quasielastic light scattering. Biochemistry. 1974 Jul 30;13(16):3336–3340. doi: 10.1021/bi00713a024. [DOI] [PubMed] [Google Scholar]
  38. Richarme G. Associative properties of the Escherichia coli galactose binding protein and maltose binding protein. Biochem Biophys Res Commun. 1982 Mar 30;105(2):476–481. doi: 10.1016/0006-291x(82)91459-0. [DOI] [PubMed] [Google Scholar]
  39. Richarme G. Interaction of the maltose-binding protein with membrane vesicles of Escherichia coli. J Bacteriol. 1982 Feb;149(2):662–667. doi: 10.1128/jb.149.2.662-667.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Saper M. A., Quiocho F. A. Leucine, isoleucine, valine-binding protein from Escherichia coli. Structure at 3.0-A resolution and location of the binding site. J Biol Chem. 1983 Sep 25;258(18):11057–11062. [PubMed] [Google Scholar]
  41. Saxton M. J. Lateral diffusion in an archipelago. Effects of impermeable patches on diffusion in a cell membrane. Biophys J. 1982 Aug;39(2):165–173. doi: 10.1016/S0006-3495(82)84504-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schindler M., Osborn M. J., Koppel D. E. Lateral mobility in reconstituted membranes--comparisons with diffusion in polymers. Nature. 1980 Jan 24;283(5745):346–350. doi: 10.1038/283346a0. [DOI] [PubMed] [Google Scholar]
  43. Shuman H. A. Active transport of maltose in Escherichia coli K12. Role of the periplasmic maltose-binding protein and evidence for a substrate recognition site in the cytoplasmic membrane. J Biol Chem. 1982 May 25;257(10):5455–5461. [PubMed] [Google Scholar]
  44. Slocum M. K., Parkinson J. S. Genetics of methyl-accepting chemotaxis proteins in Escherichia coli: organization of the tar region. J Bacteriol. 1983 Aug;155(2):565–577. doi: 10.1128/jb.155.2.565-577.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Szmelcman S., Schwartz M., Silhavy T. J., Boos W. Maltose transport in Escherichia coli K12. A comparison of transport kinetics in wild-type and lambda-resistant mutants as measured by fluorescence quenching. Eur J Biochem. 1976 May 17;65(1):13–19. doi: 10.1111/j.1432-1033.1976.tb10383.x. [DOI] [PubMed] [Google Scholar]
  46. Thompson N. L., Axelrod D. Reduced lateral mobility of a fluorescent lipid probe in cholesterol-depleted erythrocyte membrane. Biochim Biophys Acta. 1980 Mar 27;597(1):155–165. doi: 10.1016/0005-2736(80)90159-5. [DOI] [PubMed] [Google Scholar]
  47. Wandersman C., Schwartz M., Ferenci T. Escherichia coli mutants impaired in maltodextrin transport. J Bacteriol. 1979 Oct;140(1):1–13. doi: 10.1128/jb.140.1.1-13.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. van Golde L. M. Metabolism of membrane phospholipids and its relation to a novel class of oligosaccharides in Escherichia coli. Proc Natl Acad Sci U S A. 1973 May;70(5):1368–1372. doi: 10.1073/pnas.70.5.1368. [DOI] [PMC free article] [PubMed] [Google Scholar]

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