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. 1979 Oct;140(1):220–228. doi: 10.1128/jb.140.1.220-228.1979

Protein-Associated Lipid of Bacillus stearothermophilus

George L Card 1, Joan C Szuba 1, Marilyn Shimizu 1
PMCID: PMC216799  PMID: 159285

Abstract

The composition and patterns of metabolism of phospholipids isolated as part of a lipid-depleted membrane fragment (LDM fragment) and associated with the membrane adenosine triphosphatase complex have been compared with those of the bulk membrane phospholipid. The bulk lipid was extracted from washed membranes with sodium cholate. The LDM fragments, which contained a portion of the electron transport system and the membrane adenosine triphosphatase complex, were purified by chromatography with Sepharose 6B. The LDM fragment preparations contained 0.10 ± 0.02 μmol of lipid phosphorus per mg of protein, compared with 0.54 ± 0.05 μmol of lipid phosphorus per mg of protein for washed membranes. The phospholipid associated with the LDM fragments consisted of 78 ± 4% cardiolipin, 7 ± 1% phosphatidylglycerol, and 15 ± 3% phosphatidylethanolamine. Changes in the total membrane lipid composition (produced by culture conditions) did not alter the phospholipid composition of the LDM fragments. The adenosine triphosphate complex was separated from the other components of the LDM fragments by suspension of the fragments in 1% Triton X-100 and precipitation with antibody specific for the F1 component of the adenosine triphosphatase complex. The phospholipid isolated with the adenosine triphosphatase complex consisted of 86% cardiolipin, 8% phosphatidylglycerol, and 6% phosphatidylethanolamine. In pulse-chase experiments with 32P and [2-3H]glycerol, the labeling patterns of the phosphatididylglycerol and phosphatidylethanolamine associated with the LDM fragments were different from those of the bulk membrane phosphatidylglycerol and phosphatidylethanolamine. It was concluded that at least a portion of the phospholipid isolated with the LDM fragments was part of a native lipid-protein complex.

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Selected References

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

  1. Awasthi Y. C., Chuang T. F., Keenan T. W., Crane F. L. Tightly bound cardiolipin in cytochrome oxidase. Biochim Biophys Acta. 1971 Jan 12;226(1):42–52. doi: 10.1016/0005-2728(71)90176-9. [DOI] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Card G. L. Metabolism of phosphatidylglycerol, phosphatidylethanolamine, and cardiolipin of Bacillus stearothermophilus. J Bacteriol. 1973 Jun;114(3):1125–1137. doi: 10.1128/jb.114.3.1125-1137.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Downs A. J., Jones C. W. Energy conservation in Bacillus megaterium. Arch Microbiol. 1975 Oct 27;105(2):159–167. doi: 10.1007/BF00447131. [DOI] [PubMed] [Google Scholar]
  6. Griffith O. H., Jost P., Capaldi R. A., Vanderkooi G. Boundary lipid and fluid bilayer regions in cytochrome oxidase model membranes. Ann N Y Acad Sci. 1973 Dec 31;222:561–573. doi: 10.1111/j.1749-6632.1973.tb15287.x. [DOI] [PubMed] [Google Scholar]
  7. Gupta C. M., Radhakrishnan R., Khorana H. G. Glycerophospholipid synthesis: improved general method and new analogs containing photoactivable groups. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4315–4319. doi: 10.1073/pnas.74.10.4315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hachimori A., Muramatsu N., Noso Y. Studies on an ATPase of thermophilic bacteria. I. Purification and properties. Biochim Biophys Acta. 1970 Jun 10;206(3):426–437. doi: 10.1016/0005-2744(70)90158-0. [DOI] [PubMed] [Google Scholar]
  9. Helenius A., Simons K. Solubilization of membranes by detergents. Biochim Biophys Acta. 1975 Mar 25;415(1):29–79. doi: 10.1016/0304-4157(75)90016-7. [DOI] [PubMed] [Google Scholar]
  10. Jost P. C., Griffith O. H., Capaldi R. A., Vanderkooi G. Evidence for boundary lipid in membranes. Proc Natl Acad Sci U S A. 1973 Feb;70(2):480–484. doi: 10.1073/pnas.70.2.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. LESKOWITZ S., WAKSMAN B. H. Studies on immunization. 1. The effect of route of injection of bovine serum albumin in Freund adjuvant on production of circulating antibody and delayed hypersensitivity. J Immunol. 1960 Jan;84:58–72. [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Lanyi J. K. Light energy conversion in Halobacterium halobium. Microbiol Rev. 1978 Dec;42(4):682–706. doi: 10.1128/mr.42.4.682-706.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Livingston D. M. Immunoaffinity chromatography of proteins. Methods Enzymol. 1974;34:723–731. doi: 10.1016/s0076-6879(74)34094-3. [DOI] [PubMed] [Google Scholar]
  15. Marsh D., Watts A., Maschke W., Knowles P. F. Protein--immobilized lipid in dimyristoylphosphatidylcholine-substituted cytochrome oxidase: evidence for both boundary and trapped-bilayer lipid. Biochem Biophys Res Commun. 1978 Mar 30;81(2):397–402. doi: 10.1016/0006-291x(78)91546-2. [DOI] [PubMed] [Google Scholar]
  16. Mosley G. A., Card G. L., Koostra W. L. Effect of calcium and anaerobiosis on the thermostability of Bacillus stearothermophilus. Can J Microbiol. 1976 Apr;22(4):468–474. doi: 10.1139/m76-073. [DOI] [PubMed] [Google Scholar]
  17. Sandermann H., Jr Regulation of membrane enzymes by lipids. Biochim Biophys Acta. 1978 Sep 29;515(3):209–237. doi: 10.1016/0304-4157(78)90015-1. [DOI] [PubMed] [Google Scholar]
  18. Singer S. J. The molecular organization of membranes. Annu Rev Biochem. 1974;43(0):805–833. doi: 10.1146/annurev.bi.43.070174.004105. [DOI] [PubMed] [Google Scholar]
  19. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  20. Träuble H., Overath P. The structure of Escherichia coli membranes studied by fluorescence measurements of lipid phase transitions. Biochim Biophys Acta. 1973 May 25;307(3):491–512. doi: 10.1016/0005-2736(73)90296-4. [DOI] [PubMed] [Google Scholar]
  21. Vaskovsky V. E., Kostetsky E. Y. Modified spray for the detection of phospholipids on thin-layer chromatograms. J Lipid Res. 1968 May;9(3):396–396. [PubMed] [Google Scholar]
  22. Wildenauer D., Khorana H. G. The preparation of lipid-depleted bacteriorhodopsin. Biochim Biophys Acta. 1977 Apr 18;466(2):315–324. doi: 10.1016/0005-2736(77)90227-9. [DOI] [PubMed] [Google Scholar]
  23. Yoshida M., Sone N., Hirata H., Kagawa Y. A highly stable adenosine triphosphatase from a thermophillie bacterium. Purification, properties, and reconstitution. J Biol Chem. 1975 Oct 10;250(19):7910–7916. [PubMed] [Google Scholar]
  24. Yu C., Yu L., King T. E. Studies on cytochrome oxidase. Interactions of the cytochrome oxidase protein with phospholipids and cytochrome c. J Biol Chem. 1975 Feb 25;250(4):1383–1392. [PubMed] [Google Scholar]

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