1887

Abstract

Homogenates of suspensions were prepared aftergrowth in the absence and presence of chloramphenicol (500 g./ml.). Sedimentation of organelles through gradients of aqueous sucrose solutions was studied by means of a zonal centrifuge. Mitochondria (marker enzyme, malate dehydrogenase) from organisms grown with chloramphenicol sediment more slowly (median sedimentation coefficient in water at 20° = 10,600 ) than those from normal cells (median sedimentation coefficient in water at 20° = 90,000 , which confirms that the former are of smaller volume than normal mitochondria. After high-speed zonal centrifugation both mitochondria (marker enzymes, NADH and NADPH-cytochrome oxidoreductases) attain similar equilibrium densities (median density = 1·21 to 1·22). The buoyant density (median = 1·23) of peroxisomes (marker enzyme, catalase) is not altered by the inclusion of chloramphenicol in the growth medium, but the proportion of non-sedimentable catalase is increased. Chloramphenicol-inhibited growth also leads to a change in the distribution of lysosomes (marker enzymes, acid -nitrophenolphosphatase, and N-acetyl-β(D)-glucosaminidase); a marked reduction in the population at = 1·15 to 1·20 is accompanied by the appearance at = 1·09 to 1·15 of a new class of organelles containing acid hydrolases.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-67-2-161
1971-08-01
2024-12-05
Loading full text...

Full text loading...

/deliver/fulltext/micro/67/2/mic-67-2-161.html?itemId=/content/journal/micro/10.1099/00221287-67-2-161&mimeType=html&fmt=ahah

References

  1. Anderson N. G., Waters D. A., Fisher W. D., Cline G. B., Nunley C. E., Elrod L. H., Rankin C. T. 1967; Analytical techniques for cell fractions. V. Characteristics of the BXIV and BXV zonal centrifuge rotors. Analytical Biochemistry 21:235–252
    [Google Scholar]
  2. Clark-Walker G. D., Linnane A. W. 1967; The biogenesis of mitochondria in Saccharomyces cerevisiae. A comparison between cytoplasmic respiratory deficient mutant yeast and chloramphenicol inhibited wild-type cells. Journal of Cell Biology 34:1–14
    [Google Scholar]
  3. De Duve C., Berthet J., Beaufay H. 1959; Gradient centrifugation of cell particles Theory and applications. Progress in Biophysics and Biophysical Chemistry 9:326–369
    [Google Scholar]
  4. Firkin F. C., Linnane A. W. 1969; Biogenesis of mitochondria. VIII. The effect of chloramphenicol on regenerating rat liver. Experimental Cell Research 55:68–76
    [Google Scholar]
  5. Halsall H. B., Schumaker V. N. 1969; Sedimentation coefficients in the zonal centrifuge. Analytical Biochemistry 30:368–376
    [Google Scholar]
  6. Lloyd D., Brightwell R., Venables S. E., Roach G. I., Turner G. 1971; Subcellular fractionation of Tetrahymenapyriformis st by zonal centrifugation: changes in activities and distribution of enzymes during the growth cycle and on starvation. Journal of General Microbiology 65:209–223
    [Google Scholar]
  7. Lloyd D., Evans D. A., Venables S. E. 1970; The effect of chloramphenicol on growth and mitochondrial function of the flagellate Polytomella caeca . Journal of General Microbiology 61:33–41
    [Google Scholar]
  8. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
    [Google Scholar]
  9. Marchant R., Smith D. G. 1968; The effect of chloramphenicol on growth and mitochondrial structure of Pythium ultimum . Journal of General Microbiology 50:391–397
    [Google Scholar]
  10. Müller M., Baudhuin P., De Duve C. 1966; Lysosomes in Tetrahymena pyriformis . Journal of Cellular Physiology 68:165–175
    [Google Scholar]
  11. Müller M., Hogg J. F., De Duve C. 1968; Distribution of tricarboxylic acid cycle and of glyoxylate cycle enzymes between mitochondria and peroxisomes in Tetrahymena pyriformis . Journal of Biological Chemistry 243:5385–5395
    [Google Scholar]
  12. Ochoa S. 1955; Malic dehydrogenase from pig heart. Methods in Enzymology 1:735–739
    [Google Scholar]
  13. Turner G., Lloyd D. 1969; The effect of growth with chloramphenicol on the mitochondria of Tetrahymena pyriformis strain st.. Biochemical Journal 116:41 p.
    [Google Scholar]
  14. Turner G., Lloyd D. 1971; The effect of chloramphenicol on growth and mitochondrial function of the ciliate protozoon Tetrahymena pyriformis strain st.. Journal of General Microbiology 67:175–88
    [Google Scholar]
/content/journal/micro/10.1099/00221287-67-2-161
Loading
/content/journal/micro/10.1099/00221287-67-2-161
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error