Summary
The membrane system of Rhodopseudomonas capsulata, strain 37b4 is investigated in cells cultivated anaerobically in the light and in the dark, respectively, at different oxygen partial pressures (pO2). The intracytoplasmic membrane vesicles of anaerobically light grown and semiaerobically [5 mm (Hg) pO2] dark grown cells show similar diameters (30–50 nm). Growing aerobically in darkness the cells contain tubular intracytoplasmic membranes with comparable diameters. An increase of the pO2 up to 400 mm (Hg) results in a slightly decreased growth rate and in a complete inhibition of bacteriochlorophyll synthesis and intracytoplasmic membrane formation. After 8 h of cultivation under these conditions tubular membranes are still found. However, they are restricted to one cell pole only.
In order to isolate the membranes, cells were broken by means of a French pressure cell. The crude, membrane fractions (sedimented at 104000-314000xg, 60 min) are purified by centrifugation on a Ficoll density gradient. This results in the formation of three membrane fractions. The light fraction consists of small vesicular particles derived from the cytoplasmic membrane. Crude membrane fractions of all cells sedimented at 314 000xg contain a relative high percentage of these particles. Intracytoplasmic membranes of aerobically grown cells are found in a middle band. Their tubular structure remains unaffected if the cells are treated by lower pressures during homogenization. The heavy band contains the intracytoplasmic membrane vesicles from light grown and from semiaerobically dark grown cells in a highly purified form.
After treatment with butanol or NaCl and subsequently with lysozyme plus EDTA cells release flat membrane fragments which still exhibit invaginations. This shows once more that the membranes are connected to each other.
Pulse chase experiments with (2-14C)-acetate support the hypothesis that the cytoplasmic membrane and intracytoplasmic membranes are transformable into each other. So they should be looked at as a morphogenetical unit.
Zusammenfassung
Das Membransystem von Rps. capsulata setzt sich aus Cytoplasmamembran und intracytoplasmatischen Membranen zusammen. In anaeroben Lichtzellen und in Dunkelzellen unter geringen Sauerstoffpartialdrucken bestehen die intracytoplasmatischen Membranen aus Vesikeln, bei Anzucht unter hohen Sauerstoffspannungen sind sie tubulär. Auch nach einer 8stündigen Kultur bei 400 mm (Hg) Sauerstoffpartialdruck, d.h. unter Bedingungen, die eine BChl-Synthese vollständig hemmen, enthalten die Zellen noch intracytoplasmatische Tubuli an einem Zellpol.
Nach Zellaufschluß mit der French pressure cell gelang es durch anschließende fraktionierte Zentrifugation und Reinigung der Partialfraktionen über Ficoll-Gradienten 3 membranhaltige Banden zu isolieren. Die leichte Bande besteht vorwiegend aus Membranfragmenten der Cytoplasmamembran. Die mittlere Bande enthält die intracytoplasmatischen Tubuli aerob angezogener Zellen. Die schwere Bande, die den höchsten Reinheitsgrad aufweist, setzt sich aus den intracytoplasmatischen Vesikeln der Licht-bzw. der semiaeroben Dunkelzellen zusammen. 14C-Markierungsexperimente und elektronenmikroskopische Beobachtungen sprechen für morphologische und morphogenetische Zusammenhänge zwischen den Membranfraktionen und stützen damit die Hypothese, daß alle Membrantypen in einer Zelle Teile eines zusammenhängenden Membransystems sind. Die einzelnen Membrantypen können reversibel ineinander überführt werden.
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Abbreviations
- BChl:
-
Bacteriochlorophyll
- CM:
-
Cytoplasmamembran
- ICM:
-
intracytoplasmatische Membran
- pO2 :
-
Sauerstoffpartialdruck
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Lampe, H.H., Oelze, J. & Drews, G. Die Fraktionierung des Membransystems von Rhodopseudomonas capsulata und seine Morphogenese. Archiv. Mikrobiol. 83, 78–94 (1972). https://doi.org/10.1007/BF00425048
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DOI: https://doi.org/10.1007/BF00425048