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. 1976 Jan;73(1):24–28. doi: 10.1073/pnas.73.1.24

Isolation and characterization of an unsaturated fatty acid-requiring mutant of cultured mammalian cells.

T Y Chang, P R Vagelos
PMCID: PMC335831  PMID: 1750

Abstract

An unsaturated fatty acid-requiring mutant derived from Chinese hamster ovary (CHO) cells has been isolated and characterized. This mutant grows normally when oleate or other unsaturated fatty acids are supplemented in the growth medium. Unlike the wild-type CHO cells, growth stops when medium is deprived of unsaturated fatty acid. Whole cell pulse experiments with [14C]acetate or [14C]stearate indicate that the mutant is defective in unsaturated fatty acid synthesis. Enzyme assays in vitro show that the enzymatic defect of the mutant is localized to the microsomal stearoyl-CoA desaturase.

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Selected References

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

  1. Albutt E. C. A study of serum lipoproteins. J Med Lab Technol. 1966 Apr;23(2):61–82. [PubMed] [Google Scholar]
  2. Bard M., Woods R. A., Haslam J. M. Porphyrine mutants of Saccharomyces cerevisiae: correlated lesions in sterol and fatty acid biosynthesis. Biochem Biophys Res Commun. 1974 Jan 23;56(2):324–330. doi: 10.1016/0006-291x(74)90845-6. [DOI] [PubMed] [Google Scholar]
  3. Cronan J. E., Jr, Ray T. K., Vagelos P. R. Selection and characterization of an E. coli mutant defective in membrane lipid biosynthesis. Proc Natl Acad Sci U S A. 1970 Mar;65(3):737–744. doi: 10.1073/pnas.65.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hawrot E., Kennedy E. P. Biogenesis of membrane lipids: mutants of Escherichia coli with temperature-sensitive phosphatidylserine decarboxylase. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1112–1116. doi: 10.1073/pnas.72.3.1112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Jones P. D., Wakil S. J. A requirement for phospholipids by the microsomal reduced diphosphopyridine nucleotide-cytochrome c reductase. J Biol Chem. 1967 Nov 25;242(22):5267–5273. [PubMed] [Google Scholar]
  6. Karst F., Lacroute F. Isolation of pleiotropic yeast mutants requiring ergosterol for growth. Biochem Biophys Res Commun. 1973 Jun 8;52(3):741–747. doi: 10.1016/0006-291x(73)90999-6. [DOI] [PubMed] [Google Scholar]
  7. Karst F., Lacroute F. Yeast mutant requiring only a sterol as growth supplement. Biochem Biophys Res Commun. 1974 Jul 10;59(1):370–376. doi: 10.1016/s0006-291x(74)80216-0. [DOI] [PubMed] [Google Scholar]
  8. MUNKRES K. D., RICHARDS F. M. THE PURIFICATION AND PROPERTIES OF NEUROSPORA MALATE DEHYDROGENASE. Arch Biochem Biophys. 1965 Mar;109:466–479. doi: 10.1016/0003-9861(65)90391-7. [DOI] [PubMed] [Google Scholar]
  9. Machtiger N. A., Fox C. F. Biochemistry of bacterial membranes. Annu Rev Biochem. 1973;42:575–600. doi: 10.1146/annurev.bi.42.070173.003043. [DOI] [PubMed] [Google Scholar]
  10. Puck T. T., Kao F. T. Genetics of somatic mammalian cells. V. Treatment with 5-bromodeoxyuridine and visible light for isolation of nutritionally deficient mutants. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1227–1234. doi: 10.1073/pnas.58.3.1227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Raetz C. R. Isolation of Escherichia coli mutants defective in enzymes of membrane lipid synthesis. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2274–2278. doi: 10.1073/pnas.72.6.2274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rosenfeld I. S., D'Agnolo G., Vagelos P. R. Synthesis of unsaturated fatty acids and the lesion in fab B mutants. J Biol Chem. 1973 Apr 10;248(7):2452–2460. [PubMed] [Google Scholar]
  13. Silbert D. F. Genetic modification of membrane lipid. Annu Rev Biochem. 1975;44:315–339. doi: 10.1146/annurev.bi.44.070175.001531. [DOI] [PubMed] [Google Scholar]
  14. Silbert D. F., Vagelos P. R. Fatty acid mutant of E. coli lacking a beta-hydroxydecanoyl thioester dehydrase. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1579–1586. doi: 10.1073/pnas.58.4.1579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Spector A. A., Hoak J. C. An improved method for the addition of long-chain free fatty acid to protein solutions. Anal Biochem. 1969 Nov;32(2):297–302. doi: 10.1016/0003-2697(69)90089-x. [DOI] [PubMed] [Google Scholar]
  16. Strittmatter P. Protein and coenzyme interactions in the NADH-cytochrome b5 reductase system. Fed Proc. 1965 Sep-Oct;24(5):1156–1163. [PubMed] [Google Scholar]

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