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. 1974 Sep 1;62(3):860–875. doi: 10.1083/jcb.62.3.860

ELECTRON MICROSCOPE STUDY OF MITOCHONDRIAL 60S AND CYTOPLASMIC 80S RIBOSOMES FROM LOCUSTA MIGRATORIA

W Kleinow 1, W Neupert 1, F Miller 1
PMCID: PMC2109213  PMID: 4136706

Abstract

Purified mitochondrial ribosomes (60S) have been isolated from locust flight muscle. Purification could be achieved after lysis of mitochondria in 0.055 M MgCl2. Mitochondrial 60S and cytoplasmic 80S ribosomes were investigated by electron microscopy in tissue sections, in sections of pellets of isolated ribosomes, and by negative staining of ribosomal suspensions. In negatively stained preparations, mitochondrial ribosomes show dimensions of ∼270 x 210 x 215 Å; cytoplasmic ribosomes measure ∼295 x 245 x 255 Å. From these values a volume ratio of mitochondrial to cytoplasmic ribosomes of 1: 1.5 was estimated. Despite their different sedimentation constants, mitochondrial ribosomes after negative staining show a morphology similar to that of cytoplasmic ribosomes. Both types of particles show bipartite profiles which are interpreted as "frontal views" and "lateral views." In contrast to measurements on negatively stained particles, the diameter of mitochondrial ribosomes in tissue sections is ∼130 Å, while the diameter of cytoplasmic ribosomes is ∼ 180–200 Å. These data suggest a volume ratio of mitochondrial to cytoplasmic ribosomes of 1:3. Subunits of mitochondrial ribosomes (40S and 25S) were obtained by incubation under dissociating conditions before fixation in glutaraldehyde. After negative staining, mitochondrial large (40S) subunits show rounded profiles with a shallow groove on a flattened side of the profile. Mitochondrial small subunits (25S) display elongated, triangular profiles.

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Selected References

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  1. Aaij C., Nanninga N., Borst P. The structure of ribosome-like particles from rat-liver mitochondria. Biochim Biophys Acta. 1972 Aug 16;277(1):140–148. doi: 10.1016/0005-2787(72)90361-9. [DOI] [PubMed] [Google Scholar]
  2. DRAHOS V., DELONG A. A simple method for obtaining perforated supporting membranes for electron microscopy. Nature. 1960 Apr 2;186:104–104. doi: 10.1038/186104a0. [DOI] [PubMed] [Google Scholar]
  3. FRASCA J. M., PARKS V. R. A ROUTINE TECHNIQUE FOR DOUBLE-STAINING ULTRATHIN SECTIONS USING URANYL AND LEAD SALTS. J Cell Biol. 1965 Apr;25:157–161. doi: 10.1083/jcb.25.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kleinow W., Neupert W., Bücher T. Small sized ribosomes from mitochondria of locusta migratoria. FEBS Lett. 1971 Jan 12;12(3):129–133. doi: 10.1016/0014-5793(71)80050-9. [DOI] [PubMed] [Google Scholar]
  5. Kleinow W., Neupert W. RNA aus Mitochondrien des Thoraxmuskels von Locusta migratoria. Hoppe Seylers Z Physiol Chem. 1970 Oct;351(10):1205–1214. [PubMed] [Google Scholar]
  6. Kleinow W., Neupert W. The mitochondrial ribosome from Locusta migratoria: Dissociation into subunits. FEBS Lett. 1971 Jul 8;15(5):359–364. doi: 10.1016/0014-5793(71)80334-4. [DOI] [PubMed] [Google Scholar]
  7. Nanninga N. Structural aspects of ribosomes. Int Rev Cytol. 1973;35:135–188. doi: 10.1016/s0074-7696(08)60354-x. [DOI] [PubMed] [Google Scholar]
  8. Noll H. Characterization of macromolecules by constant velocity sedimentation. Nature. 1967 Jul 22;215(5099):360–363. doi: 10.1038/215360a0. [DOI] [PubMed] [Google Scholar]
  9. Nonomura Y., Blobel G., Sabatini D. Structure of liver ribosomes studied by negative staining. J Mol Biol. 1971 Sep 14;60(2):303–323. doi: 10.1016/0022-2836(71)90296-8. [DOI] [PubMed] [Google Scholar]
  10. O'Brien T. W., Kalf G. F. Ribosomes from rat liver mitochondira. II. Partial characterization. J Biol Chem. 1967 May 10;242(9):2180–2185. [PubMed] [Google Scholar]
  11. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Subramanian A. R. Glutaraldehyde fixation of ribosomes. Its use in the analysis of ribosome dissociation. Biochemistry. 1972 Jul 4;11(14):2710–2714. doi: 10.1021/bi00764a025. [DOI] [PubMed] [Google Scholar]
  13. TAHMISIAN T. N. USE OF THE FREEZING POINT METHOD TO ADJUST THE TONICITY OF FIXING SOLUTIONS. J Ultrastruct Res. 1964 Apr;10:182–188. doi: 10.1016/s0022-5320(64)80002-2. [DOI] [PubMed] [Google Scholar]
  14. Vignais P. V., Stevens B. J., Huet J., André J. Mitoribosomes from Candida utilis. Morphological, physical, and chemical characterization of the monomer form and of its subunits. J Cell Biol. 1972 Sep;54(3):468–492. doi: 10.1083/jcb.54.3.468. [DOI] [PMC free article] [PubMed] [Google Scholar]

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