Abstract
Isolation of highly purified membrane fractions from phototrophically grown Rhodospirillum rubrum was achieved by velocity and isopyknic sedimentation under carefully controlled ionic conditions. Bacteriochlorophyll-rich and succinic dehydrogenase-rich chromatophores that were essentially devoid of contamination by non-chromatophore protein were separated from a denser fraction in extracts disrupted in a French pressure cell. Highly purified chromatophores and a nearly photopigment-free envelope fraction were also obtained from cells lysed by treatment with ethylenediaminetetraacetate-lysozyme-Brij 58. After lysis with lysozyme and ethylenediaminetetraacetate alone, about 50% of the total photosynthetic pigment was released in chromatophores similar to those isolated by the above procedures. Chromatophores prepared by each method were found to have very similar near-infrared absorption spectra, overall chemical composition, equilibrium buoyant densities in CsCl, and protein patterns in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The protein profiles of the dense, outer membrane-rich fractions were different from those of the chromatophores. The release of much of the photosynthetic apparatus as discrete chromatophores is osmotically lysed extracts necessitates a reevaluation of the concept that isolated chromatophores arise only from mechanical comminution of a larger membrane structure.
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- Bayer M. E., Remsen C. C. Structure of Escherichia coli after freeze-etching. J Bacteriol. 1970 Jan;101(1):304–313. doi: 10.1128/jb.101.1.304-313.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- COHEN-BAZIRE G., KUNISAWA R. The fine structure of Rhodospirillum rubrum. J Cell Biol. 1963 Feb;16:401–419. doi: 10.1083/jcb.16.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- COHEN-BAZIRE G., SISTROM W. R., STANIER R. Y. Kinetic studies of pigment synthesis by non-sulfur purple bacteria. J Cell Physiol. 1957 Feb;49(1):25–68. doi: 10.1002/jcp.1030490104. [DOI] [PubMed] [Google Scholar]
- Cohen-Bazire G., Kunisawa R. SOME OBSERVATIONS ON THE SYNTHESIS AND FUNCTION OF THE PHOTOSYNTHETIC APPARATUS IN RHODOSPIRILLUM RUBRUM. Proc Natl Acad Sci U S A. 1960 Dec;46(12):1543–1553. doi: 10.1073/pnas.46.12.1543. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collins M. L., Niederman R. A. Membranes of Rhodospirillum rubrum: isolation and physicochemical properties of membranes from aerobically grown cells. J Bacteriol. 1976 Jun;126(3):1316–1325. doi: 10.1128/jb.126.3.1316-1325.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FRENKEL A. W., HICKMAN D. D. Structure and photochemical activity of chlorophyll-containing particles from Rhodospirillum rubrum. J Biophys Biochem Cytol. 1959 Oct;6:285–290. doi: 10.1083/jcb.6.2.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fraker P. J., Kaplan S. Isolation and fractionation of the photosynthetic membranous organelles from Rhodopseudomonas spheroides. J Bacteriol. 1971 Oct;108(1):465–473. doi: 10.1128/jb.108.1.465-473.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gibson K. D., Segen B. J., Niederman R. A. Membranes of Rhodopseudomonas spheroides. II. Precursor-product relations in anaerobically growing cells. Arch Biochem Biophys. 1972 Oct;152(2):561–568. doi: 10.1016/0003-9861(72)90251-2. [DOI] [PubMed] [Google Scholar]
- Godson G. N., Sinsheimer R. L. Lysis of Escherichia coli with a neutral detergent. Biochim Biophys Acta. 1967 Dec 19;149(2):476–488. doi: 10.1016/0005-2787(67)90175-x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Niederman R. A., Gibson K. D. The separation of chromatophores from the cell envelope in Rhodopseudomonas spheroides. Prep Biochem. 1971;1(2):141–150. doi: 10.1080/00327487108081935. [DOI] [PubMed] [Google Scholar]
- Niederman R. A. Membranes of Rhodopseudomonas spheroides: interactions of chromatophores with the cell envelope. J Bacteriol. 1974 Jan;117(1):19–28. doi: 10.1128/jb.117.1.19-28.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niederman R. A., Segen B. J., Gibson K. D. Membranes of Rhodopseudomonas spheroides. I. Isolation and characterization of membrane fractions from extracts of aerobically and anaerobically grown cells. Arch Biochem Biophys. 1972 Oct;152(2):547–560. doi: 10.1016/0003-9861(72)90250-0. [DOI] [PubMed] [Google Scholar]
- Noël H., Van der Rest M., Gingras G. Isolation and partial characterization of P870 reaction center complex from wild type Rhodospirillum rubrum. Biochim Biophys Acta. 1972 Aug 17;275(2):219–230. doi: 10.1016/0005-2728(72)90043-6. [DOI] [PubMed] [Google Scholar]
- ORMEROD J. G., ORMEROD K. S., GEST H. Light-dependent utilization of organic compounds and photoproduction of molecular hydrogen by photosynthetic bacteria; relationships with nitrogen metabolism. Arch Biochem Biophys. 1961 Sep;94:449–463. doi: 10.1016/0003-9861(61)90073-x. [DOI] [PubMed] [Google Scholar]
- Oelze J., Biedermann M., Drews G. Die Morphogenese des Photosyntheseapparates von Rhodospirillum rubrum. I. Die Isolierung und Charakterisierung von zwei Membransystemen. Biochim Biophys Acta. 1969 Apr;173(3):436–437. doi: 10.1016/0005-2736(69)90008-x. [DOI] [PubMed] [Google Scholar]
- Oelze J., Drews G. Membranes of photosynthetic bacteria. Biochim Biophys Acta. 1972 Apr 18;265(2):209–239. doi: 10.1016/0304-4157(72)90003-2. [DOI] [PubMed] [Google Scholar]
- Okamura M. Y., Steiner L. A., Feher G. Characterization of reaction centers from photosynthetic bacteria. I. Subunit structure of the protein mediating the primary photochemistry in Rhodopseudomonas spheroides R-26. Biochemistry. 1974 Mar 26;13(7):1394–1403. doi: 10.1021/bi00704a013. [DOI] [PubMed] [Google Scholar]
- SCHACHMAN H. K., PARDEE A. B., STANIER R. Y. Studies on the macro-molecular organization of microbial cells. Arch Biochem Biophys. 1952 Jul;38:245–260. doi: 10.1016/0003-9861(52)90029-5. [DOI] [PubMed] [Google Scholar]
- Salton M. R. Structure and function of bacterial cell membranes. Annu Rev Microbiol. 1967;21:417–442. doi: 10.1146/annurev.mi.21.100167.002221. [DOI] [PubMed] [Google Scholar]
- Schnaitman C. A. Examination of the protein composition of the cell envelope of Escherichia coli by polyacrylamide gel electrophoresis. J Bacteriol. 1970 Nov;104(2):882–889. doi: 10.1128/jb.104.2.882-889.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schnaitman C. A. Protein composition of the cell wall and cytoplasmic membrane of Escherichia coli. J Bacteriol. 1970 Nov;104(2):890–901. doi: 10.1128/jb.104.2.890-901.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Singer S. J., Nicolson G. L. The fluid mosaic model of the structure of cell membranes. Science. 1972 Feb 18;175(4023):720–731. doi: 10.1126/science.175.4023.720. [DOI] [PubMed] [Google Scholar]
- Takemoto J. Kinetics of photosynthetic membrane protein assembly in Rhodopseudomonas spheroides. Arch Biochem Biophys. 1974 Aug;163(2):515–520. doi: 10.1016/0003-9861(74)90509-8. [DOI] [PubMed] [Google Scholar]
- Tuttle A. L., Gest H. SUBCELLULAR PARTICULATE SYSTEMS AND THE PHOTOCHEMICAL APPARATUS OF RHODOSPIRILLUM RUBRUM. Proc Natl Acad Sci U S A. 1959 Aug;45(8):1261–1269. doi: 10.1073/pnas.45.8.1261. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- WOODY B. R., LINDSTROM E. S. The succinic dehydrogenase from Rhodospirillum rubrum. J Bacteriol. 1955 Mar;69(3):353–356. doi: 10.1128/jb.69.3.353-356.1955. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Yamashita J., Kamen M. D. Observations on distribution of NADH oxidase in particles from dark-grown and light-grown Rhodospirillum rubrum. Biochem Biophys Res Commun. 1969 Feb 21;34(4):418–425. doi: 10.1016/0006-291x(69)90398-2. [DOI] [PubMed] [Google Scholar]