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. 1971 Oct;108(1):465–473. doi: 10.1128/jb.108.1.465-473.1971

Isolation and Fractionation of the Photosynthetic Membranous Organelles from Rhodopseudomonas spheroides1

Pamela J Fraker a,2, Samuel Kaplan a
PMCID: PMC247087  PMID: 4107812

Abstract

Molecular sieve chromatography and sucrose gradient centrifugation were used to prepare large quantities of purified chromatophores from Rhodopseudomonas spheroides. Electron micrographs of these chromatophores revealed that the final preparations were very homogeneous and free of non-chromatophore particulate material. As an additional check on purity, 14C-l-phenylalanine-labeled aerobic cells, devoid of chromatophores, were mixed with unlabeled photosynthetic cells. The resulting preparation contained less than 1% of the radioactivity, originally located in non-chromatophore protein. The purified chromatophores were solubilized in 2-chloroethanol and separated into two fractions. Fraction P1 contained 3 to 5% of the total chromatophore protein and could be resolved into 10 electrophoretic components. The second fraction, PII, contained five electrophoretic components. One of these components had associated with it all of the pigment and phospholipid present in PII. Preliminary immunochemical studies on these fractions are also reported.

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Selected References

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

  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. Biedermann M., Drews G. Trennung der Thylakoidbausteine einiger Athiorhodaceae durch Gelelektrophorese. Arch Mikrobiol. 1968;61(1):48–58. [PubMed] [Google Scholar]
  3. CERIOTTI G. A microchemical determination of desoxyribonucleic acid. J Biol Chem. 1952 Sep;198(1):297–303. [PubMed] [Google Scholar]
  4. CLAYTON R. K. TOWARD THE ISOLATION OF A PHOTOCHEMICAL REACTION CENTER IN RHODOPSEUDOMONAS SPHEROIDES. Biochim Biophys Acta. 1963 Nov 29;75:312–323. doi: 10.1016/0006-3002(63)90618-8. [DOI] [PubMed] [Google Scholar]
  5. Edelhoch H. Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry. 1967 Jul;6(7):1948–1954. doi: 10.1021/bi00859a010. [DOI] [PubMed] [Google Scholar]
  6. Eytan G., Ohad I. Biogenesis of chloroplast membranes. VI. Cooperation between cytoplasmic and chloroplast ribosomes in the synthesis of photosynthetic lamellar proteins during the greening process in a mutant of Chlamydomonas reinhardi y-1. J Biol Chem. 1970 Sep 10;245(17):4297–4307. [PubMed] [Google Scholar]
  7. Gibson K. D. Electron microscopy of chromatophores of Rhodopseudomonas spheroides. J Bacteriol. 1965 Oct;90(4):1059–1072. doi: 10.1128/jb.90.4.1059-1072.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gorchein A., Neuberger A., Tait G. H. The isolation and characterization of subcellular fractions from pigmented and unpigmented cells of Rhodopseudomonas spheroides. Proc R Soc Lond B Biol Sci. 1968 Jul 2;170(1020):229–246. doi: 10.1098/rspb.1968.0035. [DOI] [PubMed] [Google Scholar]
  9. Gorchein A. The separation and identification of the lipids of Rhodopseudomonas spheroides. Proc R Soc Lond B Biol Sci. 1968 Jul 2;170(1020):279–297. doi: 10.1098/rspb.1968.0039. [DOI] [PubMed] [Google Scholar]
  10. Green D. E., Haard N. F., Lenaz G., Silman H. I. On the noncatalytic proteins of membrane systems. Proc Natl Acad Sci U S A. 1968 May;60(1):277–284. doi: 10.1073/pnas.60.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. HANDS A. R., BARTLEY W. The fatty acids of Rhodopseudomonas particles. Biochem J. 1962 Aug;84:238–239. doi: 10.1042/bj0840238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. HOLT S. C., MARR A. G. ISOLATION AND PURIFICATION OF THE INTRACYTOPLASMIC MEMBRANES OF RHODOSPIRILLUM RUBRUM. J Bacteriol. 1965 May;89:1413–1420. doi: 10.1128/jb.89.5.1413-1420.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. HOLT S. C., MARR A. G. LOCATION OF CHLOROPHYLL IN RHODOSPIRILLUM RUBRUM. J Bacteriol. 1965 May;89:1402–1412. doi: 10.1128/jb.89.5.1402-1412.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ketchum P. A., Holt S. C. Isolation and characterization of the membranes from Rhodospirillum rubrum. Biochim Biophys Acta. 1970;196(2):141–161. doi: 10.1016/0005-2736(70)90002-7. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Lockshin A., Burris R. H. Solubilization and properties of chloroplast lamellar protein. Proc Natl Acad Sci U S A. 1966 Nov;56(5):1564–1570. doi: 10.1073/pnas.56.5.1564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Marrs B., Kaplan S. 23 s precursor ribosomal RNA of Rhodopseudomonas spheroides. J Mol Biol. 1970 Apr 28;49(2):297–317. doi: 10.1016/0022-2836(70)90247-0. [DOI] [PubMed] [Google Scholar]
  18. Mazia D., Ruby A. Dissolution of erythrocyte membranes in water and comparison of the membrane protein with other structural proteins. Proc Natl Acad Sci U S A. 1968 Nov;61(3):1005–1012. doi: 10.1073/pnas.61.3.1005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Morowitz H. J., Terry T. M. Characterization of the plasma membrane of Mycoplasma laidlawii. V. Effects of selective removal of protein and lipid. Biochim Biophys Acta. 1969 Jul 15;183(2):276–294. doi: 10.1016/0005-2736(69)90084-4. [DOI] [PubMed] [Google Scholar]
  20. Oelze J., Schroeder J., Drews G. Bacteriochlorophyll, fatty-acid, and protein synthesis in relation to thylakoid formation in mutant strains of Rhodospirillum rubrum. J Bacteriol. 1970 Mar;101(3):669–674. doi: 10.1128/jb.101.3.669-674.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. SISTROM W. R. A requirement for sodium in the growth of Rhodopseudomonas spheroides. J Gen Microbiol. 1960 Jun;22:778–785. doi: 10.1099/00221287-22-3-778. [DOI] [PubMed] [Google Scholar]
  22. Schmitz R. Uber die Zusammensetzung der pigmenthaltigen Strukturen des Purpurbakteriums Rhodopseudomonas spheroides. Z Naturforsch B. 1967 Jun;22(6):645–648. [PubMed] [Google Scholar]
  23. Schnaitman C. A. Comparison of rat liver mitochondrial and microsomal membrane proteins. Proc Natl Acad Sci U S A. 1969 Jun;63(2):412–419. doi: 10.1073/pnas.63.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Takayama K., MacLennan D. H., Tzagoloff A., Stoner C. D. Studies on the electron transfer system. LXVII. Polyacrylamide gel electrophoresis of the mitochondrial electron transfer complexes. Arch Biochem Biophys. 1966 Apr;114(1):223–230. doi: 10.1016/0003-9861(66)90324-9. [DOI] [PubMed] [Google Scholar]
  25. VATTER A. E., WOLFE R. S. The structure of photosynthetic bacteria. J Bacteriol. 1958 Apr;75(4):480–488. doi: 10.1128/jb.75.4.480-488.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. WINZLER R. J. Determination of serum glycoproteins. Methods Biochem Anal. 1955;2:279–311. doi: 10.1002/9780470110188.ch10. [DOI] [PubMed] [Google Scholar]
  27. WORDEN P. B., SISTROM W. R. THE PREPARATION AND PROPERTIES OF BACTERIAL CHROMATOPHORE FRACTIONS. J Cell Biol. 1964 Oct;23:135–150. doi: 10.1083/jcb.23.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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