Abstract
-
1.
The dielectric properties of suspensions of intact cells of Methylophilus methylotrophus, Paracoccus denitrificans and Bacillus subtilis have been measured in the frequency range 1 kHz to 13 MHz. All possess a pronounced dispersion corresponding in magnitude and relaxation time to the “β-dispersion” in a terminology defined by Schwan [Adv. Biol. Med. Phys. 5: 147–209 (1957)]. The latter two strains, but not M. methylotrophus, also possess a substantial α-dispersion. The relaxation time of the β-dispersion of B. subtilis is significantly lower than that of the other two strains, due to the higher internal K+ content of this Gram-positive organism.
-
2.
Treatment of P. denitrificans or B. subtilis with lysozyme greatly reduces the magnitude of the α-dispersion; in the latter case it is virtually abolished.
-
3.
The magnitude of both the α- and β-dispersions of protoplasts of these organisms is significantly decreased by treatment with the cross-linking reagent glutaraldehyde, indicating that diffusional motions of the lipids and/or proteins in the protoplast membranes contribute to the dielectric relaxations observed in this frequency range. Such motions cannot be unrestricted, as in the “fluid mosaic” model, since the relaxation times of the lipids and proteins, if restricted by hydrodynamic forces alone, should then correspond, in protoplasts of this radius (0.4–0.5 μm), to approximately 10 Hz.
-
4.
Even after treatment of the (spherical) protoplasts with glutaraldehyde, the breadth of the remaining β-dispersion is still significantly greater than (a) that of a pure Debye dispersion and (b) that to be expected solely from a classical Maxwell-Wagner-type mechanism.
-
5.
It is recognised that the surfaces of the protein complexes in such membranes extend significantly beyond the membrane surface as delineated by the phospholipid head-groups; such molecular granularity can in principle account for the broadened dielectric relaxations in the frequency range above 1 kHz, in terms of the impediment to genuinely tangential counterion relaxation caused by the protruding proteins themselves.
-
6.
The relaxation time of a previously observed, novel, low-frequency, glutaraldehyde-sensitive (μ-) dispersion in bacterial chromatophore suspensions, as well as that of their α-dispersion, is significantly increased by increasing the aqueous viscosity with glycerol. This finding is consistent with the view that, from a dielectric standpoint, the motions of charged proteins (and lipids) in biological membranes are rather tightly coupled to those of the adjacent ions and dipoles in the electric double layer.
-
7.
Mebbrane vesicles of P. denitrificans do not possess a μ-dispersion as marked as that of chromatophores. As with chromatophores, their α-dispersion is somewhat decreased by glutaraldehyde treatment. The relative lack of a μ-dispersion in these vesicles may be related to their different polarity relative to that of bacterial chromatophores; alternatively, and perhaps additionally, the longrange lateral mobility of lipids and proteins in this system may be even more restricted than in chromatophores.
-
8.
Overall, our results draw attention to the fact that the motions of proteins, lipids and double-layer species can contribute to the audio- and radiofrequency dielectric properties of microbial cell, protoplast and vesicle suspensions, and indicate that both the magnitude and the rate of such relaxations depend rather finely on the intimate molecular structure and organisation of the bacterial cytoplasmic membrane and its superincumbent double layers.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Adey WR (1981) Tissue interactions with nonionising electromagnetic fields. Physiol Rev 61:435–514
Asami K, Irimajiri A (1984) Dielectric dispersion of a single spherical bilayer membrane in suspension. Biochim Biophys Acta 769:370–376
Asami K, Hanai T, Koizumi N (1980) Dielectric analysis of Escherichia coli in the light of the theory of interfacial polarization. Biophys J 31:215–228
Capaldi RA (1982) Arrangement of proteins in the mitochondrial inner membrane. Biochim Biophys Acta 694:291–306
Carstensen EL, Marquis RE (1975) Dielectric and electrochemical properties of bacterial cells. In: Gerhardt P, Costilow RN, Sadoff HL (eds) Spore VI. American Society for Microbiology Washington, pp 563–571
Cole KS (1972) Membranes, ions and impulses. University of California Press, Berkeley
Cole KS, Cole RH (1941) Dispersion and absorption in dielectrics. I. Alternating current chracteristics. J Chem Phys 9:341–351
Curds CR, Roberts DMcL, Wu C-H (1978) The use of continuous cultures and electronic sizing devices to study the growth rate of two species of ciliated protozoa. Soc Bacteriol Weybridge, vol 11. Academic Press, London, pp 165–177
Davies GE, Stark GR (1970) Use of dimethyl suberimidate, a cross-linking reagent, in studying the subunit structure of oligomeric proteins. Proc Natl Acad Sci USA 66:651–656
Davis DH, Doudoroff M, Stanier RY, Mandel M (1969) Proposal to reject the genus Hydrogenomonas: Taxonomic implications. Int J Syst Bacteriol 19:375–390
Donnellan EJ Jr, Nags EA, Levinson HS (1964) Chemically defined, synthetic media for sporulation and for germination and growth of Bacillus subtilis. J Bacteriol 87:332–336
Dunn G, Torgerson DM, Mandelstam J (1976) Order of expression of genes affecting septum location during sporulation of Bacillus subtilis. J Bacteriol 125:776–779
Einolf CW, Carstensen EL (1969) Passive electrical properties of microorganisms. IV. Studies of the protoplasts of Microcioccus lysodeikticus. Biophys J 9:634–643
Einolf CW, Carstensen EL (1973) Passive electrical properties of microorganisms. V. Low fequency dielectric dispersions of bacteria. Biophys J 13:8–13
Fricke H, Schwan HP, Li K, Bryson V (1956) A dielectric study of the low-conductance surface membrane in E. coli. Nature 177:134–135
Grant EH, Sheppard RJ, South GP (1978) Dielectric behaviour of biological molecules in solution. Oxford University Press, London
Grant FA (1958) Use of complex conductivity in the representation of dielectric phenomena. J Appl Phys 29:76–80
Hackenbrock CR (1981) Lateral diffusion and electron transfer in the mitochondrial inner membrane. Trends Biochem Sci 6:151–154
Harold FM (1977) Ion currents and physiological functions in microorganisms. Ann Rev Microbiol 31:181–203
Harris CM, Kell DB (1983) The radio-frequency dielectric properties of yeast cells measured with a rapid, frequencydomain dielectric spectrometer. Bioelectrochem Bioenerg 11:15–28
Harris CM, Hitchens GD, Kell DB (1984) Dielectric spectroscopy of microbial membrane systems. In: Allen MJ, Usherwood PNR (eds) Charge and field effects in biosystems. Abacus Press, Tunbridge Wells, pp 179–185
Hitchens GD, Kell DB (1982) On the extent of localization of the energised membrane state in chromatophores from Rhodopseudomonas capsulata N22. Biochem J 206:351–357
Hitchens GD, Kell DB (1984) On the effects of thiocyanate and venturicidin on respiration-driven proton translocation in Paracoccus denitrificans. Biochim Biophys Acta 766:222–232
Houslay MD, Stanley KK (1982) Dynamics of biological membranes. Wiley Chichester
John P, Whatley FR (1977) The bioenergetics of Paracoccus denitrificans. Biochim Biophys Acta 463:129–153
Junge W (1982) Electrogenic reactions and proton pumping in green plant photosynthesis. Curr Top Membr Trans 16:431–465
Kell DB (1983) Dielectric properties of bacterial chromatophores. Biolectrochem Bioenerg 11:405–415
Kell DB (1984a) Diffusion of protein complexes in prokaryotic membranes: Fast, free random or directed? Trends Biochem Sci 9:86–88
Kell DB (1984b) Dielectric spectroscopy of the rotational and translational motions of membrane proteins: Theory and experiment. EBEC Rep 3:645–646
Kell DB, Harris CM (1985a) On the dielectrically observable consequences of the diffusional motions of lipids and proteins in membranes. I. Theory and overview. Eur Biophys J 12:181–197
Kell DB, Harris CM (1985b) Dielectric spectroscopy and membrane organization. J Bioelectricity (in press)
Kell DB, Westerhoff HV (1985) Catalytic facilitation and membrane bioenergetics. In: Welch GR (ed) Organised multienzyme systems: catalytic properties. Academic Press, New York, pp 63–139
Kell DB, Ferguson SJ, John P (1978a) Determination by a flow dialysis technique of the protonmotive foree in chromatophores from Rhodospirillium rubrum. Comparison with phosphorylation potential. Biochim Biophys Acta 502:111–126
Kell DB, John P, Ferguson SJ (1978b) The protonmotive force in phosphorylating membrane vesicles from Paracoccus denitrificans. Magnitude, sites of generation and comparison with the phosphorylation potential. Biochem J 174:257–266
Kosterich JB, Foster KR, Pollack SR (1983) Dielectric permittivity and electrical conductivity of fluid-saturated bone. IEEE Trans Biomed Eng BME 30:81–86
Kubitschek HE (1969) Counting and sizing microorganisms with the Coulter counter. In: Norris JR, Ribbons DW (eds) Methods in microbiology, vol 1. Academic Press, London, pp 593–610
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Marquis RE, Carstensen EL (1973) Electric conductivity and internal osmolarity of intact bacterial cells. J Bacterial 113:1198–1206
McCarthy JEG, Ferguson SJ, Kell DB (1981) Estimation with an ion-selective electrode of the membrane potential in cells of Paracoccus denitrificans from the uptake of the butyl triphenyl phosphonium cation during aerobic and anaerobic respiration. Biochem J 196:311–321
McLaughlin SGA, Dilger JP (1980) Transport of protons across membranes by weak acids. Physiol Rev 60:825–863
Means GE, Feeney RE (1971) Chemical modification of proteins. Holden-Day San Francisco, pp 89–93
Pauly H (1962) Electrical properties of the cytoplasmic membrane and the cytoplasm of bacteria and of protoplasts. IRE Trans Biomed Electron 9:93–95
Pauly H, Packer L (1960) The relationship of internal conductance and membrane capacity to mitochondrial volume. J Biophys Biochem Cytol 7:603–612
Pauly H, Schwan HP (1959) Über die Impedanz einer Suspension von kugelförmigen Teilchen mit einer Schale. Ein Modell für das dielektrische Verhalten von Zellsuspension und von Protein Lösungen. Z Naturforsch 14 B:125–131
Pauly H, Packer L, Schwan HP (1960) Electrical properties of mitochondrial membranes. J Biophys Biochem Cytol 7:589–601
Peters K, Richards FM (1977) Chemical cross-linking reagents and problems in studies of membrane structure. Annu Rev Biochem 46:523–551
Pethig R (1979) Dielectric and electronic properties of biologyical materials, Wiley, Chichester
Pethig R (1984) Dielectric properties of biological materials: Biophysical and medical applications. IEEE Trans Electr Insulat EI19:453–474
Pilla AA (1980) Electrochemical information transfer at cell surfaces and junctions. Application to the study and manipulation of cell regulation. In: Keyzer H, Gutmann F (eds) Bioelectrochemistry. Plenum Press, New York, pp 353–396
Ramaley RF, Burden L (1970) Replacement sporulation of Bacillus subtilis 168 in a chemically defined medium. J Bacteriol 101:1–8
Rouse H (1938) Fluid mechanics for hydraulic engineers. McGraw-Hill, New York, p 407
Saffman PG, Delbrück M (1975) Brownian motion in biological membranes. Proc Natl Acad Sci USA 72:3111–3113
Schanne OF, Ceretti ERP (1978) Impedance measurements in biological cells. Wiley, Chichester
Scholes P, Smith L (1968) The isolation and properties of the cytoplasmic membrane of Micrococcus denitrificans. Biochim Biophys Acta 153:350–362
Schwan HP (1957) Electrical properties of tissue and cell suspensions. In: Lawrence JH, Tobias CA (eds) Advances in biological and medical physics, vol 5. Academic Press, New York, pp 147–209
Schwan HP (1959) Alternating current spectroscopy of biological substances. Proc IRE 47:1841–1855
Schwan HP (1977) Field interaction with biological matter. Ann NY Acad Sci 303:198–213
Schwan HP (1981a) Dielectric properties of biological tissue and biophysical mechanisms of electromagnetic field interactions. ACS Symp Ser 157:109–131
Schwan HP (1981b) Electrical properties of cells: Principles, some recent results and some unresolved problems. In: Adelmann WJ Jr, Goldman DE (eds) The biological approach to excitable systems. Plenum Press, New York, pp 3–24
Schwan HP (1983a) Dielectric properties of biological tissues and cells at RF- and MW-frequencies. In: Grandolfo M, Michaelson SM, Rindi A (eds) Biological effects and dosimetry of nonionizing radiation. Plenum Press, New York, pp 195–211
Schwan HP (1983b) Dielectric properties of biological tissues and cells at ELF-frequencies. In: Grandolfo M, Michaelson SM, Rindi A (eds) Biological effects and dosimetry of nonionizing radiation. Plenum Press, New York, pp 549–559
Schwan HP, Foster KR (1980) RF-field interactions with biological system: Electrical properties and biophysical mechanisms. Proc IEEE 68:104–113
Schwan HP, Takashima S, Miyamoto VK, Stoeckenius W (1970) Electrical properties of phospholipid vesicles. Biophys J 10:1102–1119
Smith SRL (1980) Single-cell protein. Philos Trans R Soc B 290:341–354
Stoy RD, Foster KR, Schwan HP (1982) Dielectric properties of mammalian tissue from 0.1 to 100 MHz: a summary of recent data. Phys Med Biol 27:501–513
Tempest DW (1969) Quantiative relationships between inorganic cations and anionic polymers in growing bacteria. Symp Soc Gen Microbiol 19:87–111
Vasey RB, Powell KA (1984) Single-cell protein. Biotechnol Genet Eng Rev 2:285–311
Vignais PM, Henry M-F, Sim E, Kell DB (1981) The electron transport system and hydrogenase of Paracoccus denitrificans. Curr Top Bioenerg 12:115–196
Windass JD, Worsey MJ, Pioli EM, Pioli D, Barth PT Atherton KT, Dart EC, Bryrom D, Powell K, Senior PJ (1980) Improved conversion of methanol to single-cell protein by Methylophilus methylotrophus. Nature 287:396–401
Zimmermann U,(1982) Electric field-mediated fusion and related electrical phenomena. Biochim Biophys Acta 694:227–277
Zimmermann U, Schulz J, Pilwat G (1973) Transcellular ion flow in E. coli B and electrical sizing of bacteria. Biophys J 13:1005–1012
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Harris, C.M., Kell, D.B. On the dielectrically observable consequences of the diffusional motions of lipids and proteins in membranes. Eur Biophys J 13, 11–24 (1985). https://doi.org/10.1007/BF00266305
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00266305