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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1977 Oct;74(10):4549–4551. doi: 10.1073/pnas.74.10.4549

Tritium suicide selection of mammalian cell mutants defective in the transport of neutral amino acids.

M C Finkelstein, C W Slayman, E A Adelberg
PMCID: PMC431983  PMID: 200920

Abstract

Mouse lymphocytic cells of the established line GF-14 were allowed to accumulate intracellular 3H-labeled aminoisobutyric acid (AIB), frozen, and stored over liquid N2. After internal radiation had reduced survival to 1 in 10(4), survivors were plated and tested for their ability to transport AIB. Out of 200 clones tested, two (designated GF-17 and GF-18) were found to have reductions to 13-35% of the parent in the rate of transport of AIB, L-alanine, L-proline, and L-serine; GF-18 also showed significant reductions in the rate of transport of L-glutamate and DL-cysteine. Little or no change was observed for 10 other amino acids or for thymidine. Kinetic analyses revealed that the mutants were not altered in Km for AIB uptake, but had Vmax values approximately 20% the value of the parent strain, GF-14, suggesting that either the number of AIB transport sites or the turnover rate of the sites has been reduced in the two mutants.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Breslow R. E., Goldsby R. A. Isolation and characterization of thymidine transport mutants of Chinese hamster cells. Exp Cell Res. 1969 Jun;55(3):339–346. doi: 10.1016/0014-4827(69)90567-9. [DOI] [PubMed] [Google Scholar]
  2. Burki H. J., Okada S. A comparison of the killing of cultured mammalian cells induced by decay of incorporated tritiated molecules at--196 degrees C. Biophys J. 1968 Apr;8(4):445–456. doi: 10.1016/S0006-3495(68)86499-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Christensen H. N. Some special kinetic problems of transport. Adv Enzymol Relat Areas Mol Biol. 1969;32:1–20. doi: 10.1002/9780470122778.ch1. [DOI] [PubMed] [Google Scholar]
  4. Englesberg E., Bass R., Heiser W. Inhibition of the growth of mammalian cells in cuture by amino acids and the isolation and characterization of L-phenylalanine transport. Somatic Cell Genet. 1976 Sep;2(5):411–428. doi: 10.1007/BF01542722. [DOI] [PubMed] [Google Scholar]
  5. FISCHER G. A. Studies of the culture of leukemic cells in vitro. Ann N Y Acad Sci. 1958 Dec 5;76(3):673–680. doi: 10.1111/j.1749-6632.1958.tb54884.x. [DOI] [PubMed] [Google Scholar]
  6. Farrant J., Knight S. C., McGann L. E., O'Brien J. Optimal recovery of lymphocytes and tissue culture cells following rapid cooling. Nature. 1974 May 31;249(456):452–453. doi: 10.1038/249452a0. [DOI] [PubMed] [Google Scholar]
  7. Garcia-Sancho J., Sanchez A., Christensen H. N. Role of protein dissociation in the transport of acidic amino acids by the Ehrlich ascites tumor cell. Biochim Biophys Acta. 1977 Jan 21;464(2):295–312. doi: 10.1016/0005-2736(77)90005-0. [DOI] [PubMed] [Google Scholar]
  8. HAROLD F. M., HAROLD R. L., ABRAMS A. A MUTANT OF STREPTOCOCCUS FAECALIS DEFECTIVE IN PHOSPHATE UPTAKE. J Biol Chem. 1965 Jul;240:3145–3153. [PubMed] [Google Scholar]
  9. Taub M., Englesberg E. Isolation and characterization of 5-fluorotryptophan-resistant mutants with altered L-tryptophan transport. Somatic Cell Genet. 1976 Sep;2(5):441–452. doi: 10.1007/BF01542724. [DOI] [PubMed] [Google Scholar]
  10. WERKHEISER W. C., BARTLEY W. The study of steady-state concentrations of internal solutes of mitochondria by rapid centrifugal transfer to a fixation medium. Biochem J. 1957 May;66(1):79–91. doi: 10.1042/bj0660079. [DOI] [PMC free article] [PubMed] [Google Scholar]

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