Photosynthesis Research

, Volume 116, Issue 2–3, pp 333–348 | Cite as

Membrane development in purple photosynthetic bacteria in response to alterations in light intensity and oxygen tension

  • Robert A. Niederman


Studies on membrane development in purple bacteria during adaptation to alterations in light intensity and oxygen tension are reviewed. Anoxygenic phototrophic such as the purple α-proteobacterium Rhodobacter sphaeroides have served as simple, dynamic, and experimentally accessible model organisms for studies of the photosynthetic apparatus. A major landmark in photosynthesis research, which dramatically illustrates this point, was provided by the determination of the X-ray structure of the reaction center (RC) in Blastochloris viridis (Deisenhofer and Michel, EMBO J 8:2149–2170, 1989), once it was realized that this represented the general structure for the photosystem II RC present in all oxygenic phototrophs. This seminal advance, together with a considerable body of subsequent research on the light-harvesting (LH) and electron transfer components of the photosynthetic apparatus has provided a firm basis for the current understanding of how phototrophs acclimate to alterations in light intensity and quality. Oxygenic phototrophs adapt to these changes by extensive thylakoid membrane remodeling, which results in a dramatic supramolecular reordering to assure that an appropriate flow of quinone redox species occurs within the membrane bilayer for efficient and rapid electron transfer. Despite the high level of photosynthetic unit organization in Rba. sphaeroides as observed by atomic force microscopy (AFM), fluorescence induction/relaxation measurements have demonstrated that the addition of the peripheral LH2 antenna complex in cells adapting to low-intensity illumination results in a slowing of the rate of electron transfer turnover by the RC of up to an order of magnitude. This is ascribed to constraints in quinone redox species diffusion between the RC and cytochrome bc 1 complexes arising from the increased packing density as the intracytoplasmic membrane (ICM) bilayer becomes crowded with LH2 rings. In addition to downshifts in light intensity as a paradigm for membrane development studies in Rba. sphaeroides, the lowering of oxygen tension in chemoheterotropically growing cells results in a gratuitous formation of the ICM by an extensive membrane biogenesis process. These membrane alterations in response to lowered illumination and oxygen levels in purple bacteria are under the control of a number of interrelated two-component regulatory circuits reviewed here, which act at the transcriptional level to regulate the formation of both the pigment and apoprotein components of the LH, RC, and respiratory complexes. We have performed a proteomic examination of the ICM development process in which membrane proteins have been identified that are temporally expressed both during adaptation to low light intensity and ICM formation at low aeration and are spatially localized in both growing and mature ICM regions. For these proteomic analyses, membrane growth initiation sites and mature ICM vesicles were isolated as respective upper-pigmented band (UPB) and chromatophore fractions and subjected to clear native electrophoresis for isolation of bands containing the LH2 and RC–LH1 core complexes. In chromatophores, increasing levels of LH2 polypeptides relative to those of the RC–LH1 complex were observed as ICM membrane development proceeded during light-intensity downshifts, along with a large array of other associated proteins including high spectral counts for the F1FO–ATP synthase subunits and the cytochrome bc 1 complex, as well as RSP6124, a protein of unknown function, that was correlated with increasing LH2 spectral counts. In contrast, the UPB was enriched in cytoplasmic membrane (CM) markers, including electron transfer and transport proteins, as well as general membrane protein assembly factors confirming the origin of the UPB from both peripheral respiratory membrane and sites of active CM invagination that give rise to the ICM. The changes in ICM vesicles were correlated to AFM mapping results (Adams and Hunter, Biochim Biophys Acta 1817:1616–1627, 2012), in which the increasing LH2 levels were shown to form densely packed LH2-only domains, representing the light-responsive antenna complement formed under low illumination. The advances described here could never have been envisioned when the author was first introduced in the mid-1960s to the intricacies of the photosynthetic apparatus during a lecture delivered in a graduate Biochemistry course at the University of Illinois by Govindjee, to whom this volume is dedicated on the occasion of his 80th birthday.


Light-harvesting complexes Light regulation Oxygen regulation Proteomics Reaction centers Rhodobacter sphaeroides 



Atomic force microscopy


Bacteriochlorophyll a




Cytoplasmic membrane


Fluorescence induction and relaxation


Fast repetition rate fluorescence


Intracytoplasmic membrane


Light harvesting


Core light-harvesting complex


Peripheral light-harvesting complex


Photosystem I


Photosystem II


Primary reaction center ubiquinone


Secondary reaction center ubiquinone


Photochemical reaction center


Upper-pigmented band





Work in the author’s laboratory was supported by grants from the U. S. Department of Energy (Grant No. DE-FG02-08ER15957 from the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science) and the National Science Foundation (Subaward No. 12-764). I thank Kamil Woronowicz, Raoul Frese, Oluwatobi B. Olubanjo, and Daniel Sha for their participation in these studies.


  1. Aagaard J, Sistrom WR (1972) Control of synthesis of reaction center bacteriochlorophyll in photosynthetic bacteria. Photochem Photobiol 15:209–225PubMedCrossRefGoogle Scholar
  2. Adams PG, Hunter CN (2012) Adaptation of intracytoplasmic membranes to altered light intensity in Rhodobacter sphaeroides. Biochim Biophys Acta 1817:1616–1617PubMedCrossRefGoogle Scholar
  3. Allen JP, Feher G, Yeates TO, Rees DC, Deisenhofer J, Michel H, Huber R (1986) Structural homology of reaction centers from Rhodopseudomonas sphaeroides and Rhodopseudomonas viridis as determined by X-ray diffraction. Proc Natl Acad Sci USA 83:8589–8593PubMedCrossRefGoogle Scholar
  4. Axelrod HL, Okamura M (2005) The structure and function of the cytochrome c 2: reaction center electron transfer complex from Rhodobacter sphaeroides. Photosynth Res 85:101–114PubMedCrossRefGoogle Scholar
  5. Bahatyrova S, Frese RN, Siebert CA, van der Werf KO, van Grondelle R, Niederman RA, Bullough PA, Otto C, Olsen JD, Hunter CN (2004) The native architecture of a photosynthetic membrane. Nature 430:1058–1062PubMedCrossRefGoogle Scholar
  6. Bauer C, Setterdahl A, Wu J, Robinson B (2008) Regulation of gene expression in response to oxygen tension. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 707–725Google Scholar
  7. Bowman WC, Du S, Bauer CE, Kranz RG (1999) In vitro activation and repression of photosynthesis gene transcription in Rhodobacter capsulatus. Mol Microbiol 33:429–437PubMedCrossRefGoogle Scholar
  8. Bowyer JR, Hunter CN, Ohnishi T, Niederman RA (1985) Photosynthetic membrane development in Rhodopseudomonas sphaeroides: spectral and kinetic characterization of redox components of light-driven electron flow in apparent photosynthetic membrane growth initiation sites. J Biol Chem 260:3295–3304PubMedGoogle Scholar
  9. Braatsch S, Gomelsky M, Kuphal S, Klug G (2002) A single flavoprotein, AppA, integrates both redox and light signals in Rhodobacter sphaeroides. Mol Microbiol 45:827–836PubMedCrossRefGoogle Scholar
  10. Braatsch S, Johnson JA, Noll K, Beatty JT (2007) The O2-responsive repressor PpsR2 but not PpsR1 transduces a light signal sensed by the BphP1 phytochrome in Rhodopseudomonas palustris CGA009. FEMS Microbiol Lett 272:60–64PubMedCrossRefGoogle Scholar
  11. Chang CH, Tiede D, Tang J, Smith U, Norris J, Schiffer M (1986) Structure of Rhodopseudomonas sphaeroides R-26 reaction center. FEBS Lett 205:82–86PubMedCrossRefGoogle Scholar
  12. Chuartzman SG, Nevo R, Shimoni E, Charuvi D, Kiss V, Ohad I, Brumfeld V, Reich Z (2008) Thylakoid membrane remodeling during state transitions in Arabidopsis. Plant Cell 20:1029–1039PubMedCrossRefGoogle Scholar
  13. Cogdell RJ, Howard TD, Bittl R, Schlodder E, Geisenheimer I, Lubitz W (2000) How carotenoids protect bacterial photosynthesis. Philos Trans R Soc Lond B 355:1345–1349CrossRefGoogle Scholar
  14. Cohen-Bazire G, Sistrom WR, Stanier RY (1957) Kinetic studies of pigment synthesis by non-sulfur purple bacteria. J Cell Comp Physiol 49:25–68CrossRefGoogle Scholar
  15. Deisenhofer J, Michel M (1989) Nobel lecture: the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis. EMBO J 8:2149–2170PubMedGoogle Scholar
  16. Drews G, Niederman RA (2002) Membrane biogenesis in anoxygenic photosynthetic prokaryotes. Photosynth Res 73:87–94PubMedCrossRefGoogle Scholar
  17. Eraso JM, Kaplan S (1994) prrA, a putative response regulator involved in oxygen regulation of photosynthesis gene expression in Rhodobacter sphaeroides. J Bacteriol 176:32–43PubMedGoogle Scholar
  18. Evans K, Fordham-Skelton AP, Mistry H, Reynolds CD, Lawless AM, Papiz MZ (2005) A bacteriophytochrome regulates the synthesis of LH4 complexes in Rhodopseudomonas palustris. Photosynth Res 85:169–180PubMedCrossRefGoogle Scholar
  19. Evans K, Georgiou T, Hillon T, Fordham-Skelton A, Papiz M (2008) Bacteriophytochromes control photosynthesis in Rhodopseudomonas palustris. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 799–809Google Scholar
  20. Feniouk BA, Junge W (2008) Proton translocation and ATP synthesis by the FOF1-ATPase of purple bacteria. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 475–493Google Scholar
  21. Francia F, Dezi M, Rebecchi A, Mallardi A, Palazzo G, Melandri BA, Venturoli G (2004) Light harvesting complex 1 stabilizes P+QB charge separation in reaction centers of Rhodobacter sphaeroides. Biochemistry 43:14199–14210PubMedCrossRefGoogle Scholar
  22. Giraud E, Fardoux J, Fourrier N, Hannibal L, Genty B, Bouyer P, Dreyfus B, Verméglio A (2002) Bacteriophytochrome controls photosystem synthesis in anoxygenic bacteria. Nature 417:202–205Google Scholar
  23. Giraud E, Zappa S, Vuillet L, Adriano JM, Hannibal L, Fardoux J, Berthomieu C, Bouyer P, Pignol D, Verméglio A (2005) A new type of bacteriophytochrome acts in tandem with a classical bacteriophytochrome to control the antennae synthesis in Rhodopseudomonas palustris. J Biol Chem 280:32389–32397PubMedCrossRefGoogle Scholar
  24. Gomelsky M, Kaplan S (1995a) Genetic evidence that PpsR from Rhodobacter sphaeroides 2.4.1 functions as a repressor of puc and bchF expression. J Bacteriol 177:1634–1637PubMedGoogle Scholar
  25. Gomelsky M, Kaplan S (1995b) AppA, a novel gene encoding a trans-acting factor involved in the regulation of photosynthesis gene expression in Rhodobacter sphaeroides 2.4.1. J Bacteriol 177:4609–4618PubMedGoogle Scholar
  26. Gomelsky M, Klug G (2002) BLUF: a novel FAD-binding domain involved in sensory transduction in microorganisms. Trends Biochem Sci 27:497–500PubMedCrossRefGoogle Scholar
  27. Hunter CN, vanGrondelle R, Holmes NG, Jones OTG, Niederman RA (1979) Fluorescence yield properties of a fraction enriched in newly synthesized bacteriochlorophyll a-protein complexes from Rhodopseudomonas sphaeroides. Photochem Photobiol 30:313–316PubMedCrossRefGoogle Scholar
  28. Hunter CN, Pennoyer JD, Sturgis JN, Farrelly D, Niederman RA (1988) Oligomerization states and associations of light-harvesting pigment–protein complexes of Rhodobacter sphaeroides as analyzed by lithium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochemistry 27:3459–3467CrossRefGoogle Scholar
  29. Hunter CN, Tucker JD, Niederman RA (2005) The assembly and organisation of photosynthetic membranes in Rhodobacter sphaeroides. Photochem Photobiol Sci 4:1023–1027PubMedCrossRefGoogle Scholar
  30. Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) (2008) The purple phototrophic bacteria. Advances in photosynthesis and respiration, vol 28. Springer, DordrechtGoogle Scholar
  31. Kim SK, Mason JT, Knaff DB, Bauer CE, Setterdahl AT (2006) Redox properties of the Rhodobacter sphaeroides transcriptional regulatory proteins PpsR and AppA. Photosynth Res 89:89–98PubMedCrossRefGoogle Scholar
  32. Kirchhoff H, Haase W, Wegner S, Danielsson R, Ackermann R, Albertsson PA (2007) Low-light-induced formation of semicrystalline photosystem II arrays in higher plant chloroplasts. Biochemistry 46:11169–11176PubMedCrossRefGoogle Scholar
  33. Klug G, Masuda S (2008) Regulation of genes by light. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 727–741Google Scholar
  34. Koblízek M, Shih JD, Breitbart SI, Ratcliffe EC, Kolber ZS, Hunter CN, Niederman RA (2005) Sequential assembly of photosynthetic units in Rhodobacter sphaeroides as revealed by fast repetition rate analysis of variable bacteriochlorophyll a fluorescence. Biochim Biophys Acta 1706:220–231PubMedCrossRefGoogle Scholar
  35. Lavergne J, Verméglio A, Joliot P (2008) Functional coupling between reaction centers and cytochrome bc 1 complexes. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 509–536Google Scholar
  36. Mank NN, Berghoff BA, Hermanns YN, Klug G (2012) Regulation of bacterial photosynthesis genes by the small noncoding RNA PcrZ. Proc Natl Acad Sci USA 109:16306–16311PubMedCrossRefGoogle Scholar
  37. Masuda S, Bauer CE (2002) AppA is a blue light photoreceptor that antirepresses photosynthesis gene expression in Rhodobacter sphaeroides. Cell 110:613–623PubMedCrossRefGoogle Scholar
  38. Metz S, Haberzettl K, Frühwirth S, Teich K, Hasewinkel C, Klug G (2012) Interaction of two photoreceptors in the regulation of bacterial photosynthesis genes. Nucleic Acids Res 40:5901–5909PubMedCrossRefGoogle Scholar
  39. Monger TG, Cogdell RJ, Parson WW (1976) Triplet states of bacteriochlorophyll and carotenoids in chromatophores of photosynthetic bacteria. Biochim Biophys Acta 449:136–153PubMedCrossRefGoogle Scholar
  40. Niederman RA (2006) Structure, function and formation of bacterial intracytoplasmic membranes. In: Shively JM (ed) Complex intracellular structures in prokaryotes, microbiology monographs, vol 2. Springer, Berlin, pp 193–227CrossRefGoogle Scholar
  41. Niederman RA, Mallon DE, Langan JJ (1976) Membranes of Rhodopseudomonas sphaeroides. IV. Assembly of chromatophores in low-aeration cell suspensions. Biochim Biophys Acta 440:429–447PubMedCrossRefGoogle Scholar
  42. Niederman RA, Mallon DE, Parks LC (1979) Membranes of Rhodopseudomonas sphaeroides. VI. Isolation of a fraction enriched in newly synthesized bacteriochlorophyll a-protein complexes. Biochim Biophys Acta 555:210–220PubMedCrossRefGoogle Scholar
  43. Papiz MZ, Prince SM, Howard T, Cogdell RJ, Isaacs NW (2003) The structure and thermal motion of the B800–850 LH2 complex from Rhodopseudomonas acidophila at 2.0 Å resolution and 100 K: new structural features and functionally relevant motions. J Mol Biol 326:1523–1538PubMedCrossRefGoogle Scholar
  44. Parson WW, Warshel A (2008) Mechanism of charge separation in purple bacterial reaction centers. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 355–377Google Scholar
  45. Ponnampalam SN, Bauer CE (1997) DNA binding characteristics of CrtJ. A redox-responding repressor of bacteriochlorophyll, carotenoid, and light harvesting-II gene expression in Rhodobacter capsulatus. J Biol Chem 272:18391–18396PubMedCrossRefGoogle Scholar
  46. Qian P, Hunter CN, Bullough PA (2005) The 8.5 Å projection structure of the core RC-LH1-PufX dimer of Rhodobacter sphaeroides. J Mol Biol 349:948–960PubMedCrossRefGoogle Scholar
  47. Reilly PA, Niederman RA (1986) Role of apparent membrane growth initiation sites during photosynthetic membrane development in synchronously dividing Rhodopseudomonas sphaeroides. J Bacteriol 167:153–159PubMedGoogle Scholar
  48. Rhee K-H, Morris EP, Barber J, Kühlbrandt W (1998) Three-dimensional structure of the plant photosystem II reaction centre at 8 Å resolution. Nature 396:283–286Google Scholar
  49. Rochaix JD (2007) Role of thylakoid protein kinases in photosynthetic acclimation. FEBS Lett 581:2768–2775PubMedCrossRefGoogle Scholar
  50. Roszak AW, Howard TD, Southall J, Gardiner AT, Law CJ, Isaacs NW, Cogdell RJ (2003) Crystal structure of the RC-LH1 core complex from Rhodopseudomonas palustris. Science 302:1969–1972PubMedCrossRefGoogle Scholar
  51. Schumacher A, Drews G (1979) Effects of light intensity on membrane differentiation in Rhodopseudomonas capsulata. Biochim Biophys Acta 547:417–428PubMedCrossRefGoogle Scholar
  52. Sturgis JN, Niederman RA (2008a) Atomic force microscopy reveals multiple patterns of antenna organization in purple bacteria: implications for energy transduction mechanisms and membrane modeling. Photosynth Res 95:269–278PubMedCrossRefGoogle Scholar
  53. Sturgis JN, Niederman RA (2008b) Organization and assembly of light-harvesting complexes in the purple bacterial membrane. In: Hunter CN, Daldal F, Thurnauer MC, Beatty JT (eds) The purple phototrophic bacteria. Springer, Dordrecht, pp 253–273Google Scholar
  54. Sturgis JN, Hunter CN, Niederman RA (1988) Spectra and extinction coefficients of near-infrared absorption bands in membranes of Rhodobacter sphaeroides mutants lacking light-harvesting and reaction center complexes. Photochem Photobiol 48:243–247CrossRefGoogle Scholar
  55. Sturgis JN, Tucker JD, Olsen JD, Hunter CN, Niederman RA (2009) Atomic force microscopy studies of native photosynthetic membranes. Biochemistry 48:3679–3698PubMedCrossRefGoogle Scholar
  56. Swem LR, Bauer CE (2002) Coordination of ubiquinol oxidase and cytochrome ccb 3 oxidase expression by multiple regulators. J Bacteriol 184:2815–2820PubMedCrossRefGoogle Scholar
  57. Swem LR, Kraft BJ, Swem DL, Setterdahl AT, Masuda S, Knaff DB, Zaleski JM, Bauer CE (2003) Signal transduction by the global regulator RegB is mediated by a redox-active cysteine. EMBO J 22:4699–4708PubMedCrossRefGoogle Scholar
  58. Swem LR, Gong X, Yu CA, Bauer CE (2006) Identification of a ubiquinone-binding site that affects autophosphorylation of the sensor kinase RegB. J Biol Chem 281:6768–6775PubMedCrossRefGoogle Scholar
  59. Tavano CL, Donohue TJ (2006) Development of the bacterial photosynthetic apparatus. Curr Opin Microbiol 9:625–631PubMedCrossRefGoogle Scholar
  60. Tucker JD, Siebert CA, Escalante M, Adams PG, Olsen JD, Otto C, Stokes DL, Hunter CN (2010) Membrane invagination in Rhodobacter sphaeroides is initiated at curved regions of the cytoplasmic membrane, then forms both budded and fully detached spherical vesicles. Mol Microbiol 76:833–847PubMedCrossRefGoogle Scholar
  61. Vredenberg WJ, Duysens LN (1963) Transfer of energy from bacteriochlorophyll to a reaction centre during bacterial photosynthesis. Nature 197:355–357PubMedCrossRefGoogle Scholar
  62. Woronowicz K, Niederman RA (2010) Proteomic analysis of the developing intracytoplasmic membrane in Rhodobacter sphaeroides during adaptation to low light intensity. Adv Exp Med Biol 675:161–178PubMedCrossRefGoogle Scholar
  63. Woronowicz K, Sha D, Frese RN, Niederman RA (2011a) The accumulation of the light-harvesting 2 complex during remodeling of the Rhodobacter sphaeroides intracytoplasmic membrane results in a slowing of the electron transfer turnover rate of photochemical reaction centers. Biochemistry 50:4819–4829PubMedCrossRefGoogle Scholar
  64. Woronowicz K, Sha D, Frese RN, Sturgis JN, Nanda V, Niederman RA (2011b) The effects of protein crowding in bacterial photosynthetic membranes on the flow of quinone redox species between the photochemical reaction center and the ubiquinol-cytochrome c 2 oxidoreductase. Metallomics 3:765–774PubMedCrossRefGoogle Scholar
  65. Woronowicz K, Olubanjo OB, Sung HC, Lamptey J, Niederman RA (2012) Differential assembly of polypeptides of the light-harvesting 2 complex encoded by distinct operons during acclimation of Rhodobacter sphaeroides to low light intensity. Photosynth Res 111:125–138PubMedCrossRefGoogle Scholar
  66. Woronowicz K, Harrold JW, Kay JM, Niederman RA (2013) Structural and functional proteomics of intracytoplasmic membrane assembly in Rhodobacter sphaeroides. J Mol Microbiol Biotechnol 23(1–2):48–62PubMedCrossRefGoogle Scholar
  67. Zeilstra-Ryalls JH, Kaplan S (1995) Aerobic and anaerobic regulation in Rhodobacter sphaeroides 2.4.1: the role of the fnrL gene. J Bacteriol 177:6422–6431PubMedGoogle Scholar
  68. Zeng X, Choudhary M, Kaplan S (2003) A second and unusual pucBA operon of Rhodobacter sphaeroides 2.4.1: genetics and function of the encoded polypeptides. J Bacteriol 185:6171–6184PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.Department of Molecular Biology and BiochemistryRutgers UniversityPiscatawayUSA

Personalised recommendations