Application of laser beat frequency spectroscopy to the detection and characterization of an oncogenic RNA virus

Salmeen, Irving T.; Gill, D.; Rimai, L.; McCormick, J. Justin; Maher, Veronica M.; Arnold, W; Hight, M.E.

doi:10.1016/0006-291x(72)90958-8

Cited by 9 publications

(4 citation statements)

References 3 publications

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“…This time dependence is due to the Brownian motion of particles in solution and, for a monodisperse suspension of spherical particles, can be analyzed to yield the diffusion constant and an estimate of the particle concentration (Schaefer and Berne, 1972). Laser beat frequency spectroscopy has been used previously for the study of proteins, viruses, bacteriophages, red blood cells, and bacterial motility (Dubin et al, 1971;Salmeen et al, 1972;Camerini-Otero et al, 1974).…”

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Hydrodynamic diameters of RNA tumor viruses. Studies by laser beat frequency light scattering spectroscopy of avian myeloblastosis and Rauscher murine leukemia viruses

Salmeen¹,

Rimai²,

Liebes³

et al. 1975

Biochemistry

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The diffusion constants of avian myeloblastosis virus (AMV) and murine leukemia virus (MuLV) (Rauscher) suspensions in buffer and in 30% sucrose were determined by laser beat frequency light scattering spectroscopy at a series of temperatures ranging rom 5 to 25 degrees. By the use of the Stokes-Einstein equation, the following hydrodynamic diameters are calculated at 20 degrees: MuLV, 154 plus or minus 3 nm in sucrose and 145 plus or minus 7 nm in buffer; AMV, 144 plus or minus 3 nm in sucrose and 138 plus or minus 4 nm in buffer. While the diameters measured in buffer were temperature independent, the diameters measured in sucrose decreased by about 20% as the temperature was raised from 5 to 25 degrees. The concentration of virus particles in the suspensions ranged from 10 7 to 10 9 particles/ml. The absolute particle concentrations are estimated within plus or minus 30% by determining the dilution needed to reach a concentration sufficiently low that the particle number fluctuation contribution was comparable to that of the interference scattering. Particle weights of 3.9 x 10 8 daltons for MuLV and 4.0 x 10 8 daltons for AMV were calculated from the diffusion constants and from our own experimentally determined sedimentation coefficients. From these particle weights and the hydrodynamic diameters of the viruses, we calculated the per cent of the hydrodynamic volume of the viruses which could be freely penetrated by water, viz., 57% for AMV and 69% for MuLV.

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Hydrodynamic diameters of RNA tumor viruses. Studies by laser beat frequency light scattering spectroscopy of avian myeloblastosis and Rauscher murine leukemia viruses

Salmeen¹,

Rimai²,

Liebes³

et al. 1975

Biochemistry

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“…However, preliminary studies (R. Luftig and K. Culbreth, unpublished observations) suggest that the smaller diameter seen in freeze-dried preparations (24) may be due to shrinkage of the virions during preparation. The recent report (29)…”

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confidence: 99%

Physicochemical Studies on L-Cell Virions

Nichols

Quade

Luftig

1973

J Virol

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The L-cell virion (LCV) has been purified from supernatant fluids of mouse L cells grown in suspension culture. The virion is similar to other RNA tumor viruses by several criteria: (i) the density of the virion is 1.16 g/cm 3 ; (ii) the virion appears as a rounded membranous particle with an outer diameter of 146.7 ± 11.8 nm, and contains knobs (7-nm diameter) over its surface; (iii) 15 polypeptides (ranging in molecular weight from 7,000 to 110,000) are detectable after electrophoresis of virion protein in sodium dodecyl sulfate-polyacrylamide gels; (iv) three species of RNA can be isolated—high molecular weight (80 to 88 s ) (50%), 7 s (35%) and 4 s (15%); (v) heat denaturation of the high-molecular-weight RNA yields a heterogeneous population of molecules (20 to 35 s ) as well as a 7 s and 4 s species. Despite the general similarity to infectious RNA tumor viruses, LCV is apparently defective as evidenced by the fact that it does not induce tumors in animals or transform normal mouse cells in vitro (Kindig and Kirsten [17]). The defective nature of the LCV might be related to the fact that assays for DNA polymerase in the virion showed only a negligible activity when compared to Rous sarcoma virus.

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“…Presently, the only way to solve this problem is by electron-microscopy. However, recent developments in the field of quasi elastic light scattering techniques give a possibility, immediately after preparation, to describe the homogeneity and to study possible structural changes in the membrane structure.Quasi elastic laser light scattering (QELS) [1,28] has been already used to determine in aqueous medium, the size of large molecular organizations: for instance, viruses [6,24,25], muscular proteins [11], and so on. The purpose of the present paper is to study the possibilities offered by quasi 7 J. Membrane Biol.…”

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Description by quasi elastic laser light scattering of a biological preparation: Sarcoplasmic reticulum vesicles

et al. 1974

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Summary. Quasi elastic laser light scattering (QELS) was used to describe the size and the homogeneity of a membrane preparation: vesicles from skeletal sarcoplasmic reticulum. The data were compared with results obtained by electron-microscopy. The advantages of each method are discussed. By electron-microscopy the average value for the diameter of the vesicles obtained after negative staining is 0.100-t-0.040 ltm. With the freeze-etching technique this value ranges from 0.13 to 0.25 ~tm. For the same biological preparation the analysis of the QELS recording curves displaying one correlation time (r) gives an apparent diameter for the vesicles of 0.17 lam. Although it is difficult to describe by QELS the homogeneity of the preparation, the technique remains a very convenient method for evaluating variation of the size of the vesicles, and thus offers the possibility to follow general modification in the size of a vesicular preparation. The effect of trypsin and phospholipase C treatment on vesicle size is studied: an increase of vesicle size is clearly observed after trypsin treatment.A direct evaluation of the homogeneity and the size of constituents of a membrane preparation after extraction and purification is a major problem. Presently, the only way to solve this problem is by electron-microscopy. However, recent developments in the field of quasi elastic light scattering techniques give a possibility, immediately after preparation, to describe the homogeneity and to study possible structural changes in the membrane structure.Quasi elastic laser light scattering (QELS) [1,28] has been already used to determine in aqueous medium, the size of large molecular organizations: for instance, viruses [6,24,25], muscular proteins [11], and so on. The purpose of the present paper is to study the possibilities offered by quasi 7 J. Membrane Biol. 18

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Application of laser beat frequency spectroscopy to the detection and characterization of an oncogenic RNA virus

Cited by 9 publications

References 3 publications

Hydrodynamic diameters of RNA tumor viruses. Studies by laser beat frequency light scattering spectroscopy of avian myeloblastosis and Rauscher murine leukemia viruses

Hydrodynamic diameters of RNA tumor viruses. Studies by laser beat frequency light scattering spectroscopy of avian myeloblastosis and Rauscher murine leukemia viruses

Physicochemical Studies on L-Cell Virions

Description by quasi elastic laser light scattering of a biological preparation: Sarcoplasmic reticulum vesicles

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