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Probing the structure of the core light-harvesting complex (LH1) of Rhodopseudomonas viridis by dissociation and reconstitution methodology

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Abstract

A subunit complex was formed from the core light-harvesting complex (LH1) of bacteriochlorophyll(BChl)-b-containing Rhodopseudomonas viridis. The addition of octyl glucoside to a carotenoid-depleted Rps. viridis membrane preparation resulted in a subunit complex absorbing at 895 nm, which could be quantitatively dissociated to free BChl b and then reassociated to the subunit. When carotenoid was added back, the subunit could be reassociated to LH1 with a 25% yield. Additionally, the Rps. viridis α- and β-polypeptides were isolated, purified, and then reconstituted with BChl b. They formed a subunit absorbing near 895 nm, similar to the subunit formed by titration of the carotenoid depleted membrane, but did not form an LH1-type complex at 1015 nm. The same results were obtained with the β-polypeptide alone and BChl b. Isolated polypeptides were also tested for their interaction with BChl a. They formed subunit and LH1-type complexes similar to those formed using polypeptides isolated from BChl-a-containing bacteria but displayed 6–10 nm smaller red shifts in their long-wavelength absorption maxima. Thus, the larger red shift of BChl-b-containing Rps. viridis is not attributable solely to the protein structure. The β-polypeptide of Rps. viridis differed from the other β-polypeptides tested in that it could form an LH1-type complex with BChl a in the absence of the α- and γ-polypeptides. It apparently contains the necessary information required to assemble into an LH1-type complex. When the γ-polypeptide was tested in reconstitution with BChl a and BChl b with the α- and β-polypeptides, it had no effect; its role remains undetermined.

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Abbreviations

B820:

the subunit form of the core light-harvesting complex in BChl-a-containing bacteria which has an absorption maximum at or near 820 nm

B875:

the core light-harvesting complex of Rhodobacter sphaeroides which has an absorption maximum at 875 nm

B881:

the core light-harvesting complex of wild-type Rhodospirillum rubrum which has an absorption maximum at 881 nm

B895:

the subunit form of the core light-harvesting complex in Rps. viridis which has an absorption maximum near 888–895 nm

B1015:

the core light-harvesting complex of Rps. viridis which has an absorption maximum at 1015 nm

CD:

circular dichroism

LH1:

the core light-harvesting complex

OG:

n-octyl β-d-glucopyranoside

References

  • Braun P and Scherz A (1991) Polypeptides and bacteriochlorophyll organization in the light-harvesting complex B850 of Rhodobacter sphaeroides R-26.1. Biochemistry 30: 5177–5184

    PubMed  Google Scholar 

  • Broglie RM, Hunter CN, Delepelaire P, Niederman RA, Chua NH and Clayton RK (1980) Isolation and characterization of the pigment-protein complexes of Rhodopseudomonas sphaeroides by lithium dodecyl sulfate/polyacrylamide gel electrophoresis. Proc Natl Acad Sci USA 77: 87–91

    PubMed  Google Scholar 

  • Brunisholz RA and Zuber H (1988) Primary structure analyses of bacterial antenna polypeptides: correlation of aromatic amino acids with spectral properties. Structural similarities with reaction center polypeptides. In: Scheer H and Schneider S (eds) Photosynthetic Light-Harvesting Systems, pp 103–114. Walter de Gruyter & Co, New York

    Google Scholar 

  • Brunisholz RA and Zuber H (1992) Structure, function and organization of antenna polypeptides and antenna complexes from the three families of Rhodospirillaneae. J Photochem Photobiol B: Biol 15: 113–140

    Article  Google Scholar 

  • Brunisholz RA, Jay F, Suter F and Zuber H (1985) The light-harvesting polypeptides of Rhodopseudomonas viridis: The complete amino-acid sequences of B1015-α, B1015-β and B1015-γ. Biol. Chem. Hoppe-Seyler 366: 87–98

    PubMed  Google Scholar 

  • Brunisholz RA, Steiner R, Scheer H, Imhoff JF and Zuber H (1991) Small amphipathic polypeptides: The decisive structural elements for far-red shifted bacteriochlorophyll-b containing membrane-bound antenna complexes? Experientia 47: A71

  • Callahan PM, Cotton T and Loach PA (1987) Borohydride reduction of bacteriochlorophyll a in the light harvesting protein of Rhodospirillum rubrum. In: Biggins J (ed) Progress in Photosinthesis Research, pp I.3.325-I.3.328. Nijhoff, Dordrecht, The Netherlands

    Google Scholar 

  • Chang MC, Callahan PM, Parkes-Loach PS, Cotton TM and Loach PA (1990a) Spectroscopic characterization of the light-harvesting complex of Rhodospirillum rubrum and its structural subunit. Biochemistry 29: 421–429

    PubMed  Google Scholar 

  • Chang MC, Meyer L and Loach PA (1990b) Isolation and characterization of a structural subunit of the core light-harvesting complex of Rhodobacter sphaeroides 2.4.1 and puc705-BA. Photochem Photobiol 52: 873–881

    PubMed  Google Scholar 

  • Cogdell RJ (1986) Carotenoid-bacteriochlorophyll interactions. Springer Ser Chem Phys 42: 62–66

    Google Scholar 

  • Cogdell RJ (1986) Light-harvesting complexes in the purple photosynthetic bacteria. Encycl Plant Physiol, New Ser 19: 252–259

    Google Scholar 

  • Cogdell RJ and Thornber JP (1979) The preparation and characterization of different types of light-harvesting complexes from some purple bacteria. Ciba Found Symp 61 (new ser): 61–79

    Google Scholar 

  • Cohen-Bazire G, Sistrom WR and Stanier RY (1957) Kinetic studies of pigment synthesis by non-sulfur purple bacteria. J Cell Comp Physiol 49: 25–68

    Google Scholar 

  • Drews G (1985) Structure and functional organization of light-harvesting complexes and photochemical reaction centers in membranes of phototrophic bacteria. Microbiol Rev 49: 59–70

    PubMed  Google Scholar 

  • Feick R and Drews G (1978) Isolation and characterization of light-harvesting bacteriochlorophyll-protein complexes from Rhodopseudomonas capsulata. Biochim Biophys Acta 501: 499–513

    PubMed  Google Scholar 

  • Fowler GJS, Visschers RW, Grief GG, van Grondelle R and Hunter CN (1992) Genetically modified photosynthetic antenna complexes with blueshifted absorbance bands. Nature (London) 355: 848–850

    Google Scholar 

  • Garcia A, Vernon LP, Ke B and Mollenhauer H (1968) Some structural and photochemical properties of Rhodopseudomonas species NHTC 133 subchromatophore particles obtained by treatment with Triton X-100. Biochemistry 7: 326–332

    PubMed  Google Scholar 

  • Ghosh R, Kessi J, Hauser H, Wehrli E and Bachofen R (1990) Quaternary structure of the B875 light-harvesting complex from Rhodospirillum rubrum G9+. FEMS Symp 53 (Mol Biol Membr-Bound Complexes Phototrophic Bact): 245–251

    Google Scholar 

  • Giesbrecht P and Drews G (1966) Über die organisation und die makromolekuläre architektur der thylakoide ‘lebender’ bakterien. Arch Mikrobiol 54: 297–330

    Google Scholar 

  • Gottstein J and Scheer H (1983) Long-wavelength-absorbing forms of bacteriochlorophyll a in solutions of Triton X-100. Proc Natl Acad Sci USA 80: 2231–2234

    Google Scholar 

  • Hawthornthwaite AM and Cogdell RJ (1991) Bacteriochlorophyllbinding proteins. In: Scheer H (ed) Chlorophylls, pp 493–528. CRC Press, Boca Raton, FL

    Google Scholar 

  • Heller BA and Loach PA (1990) Isolation and characterization of a subunit form of the B875 light-harvesting complex from Rhodobacter capsulatus. Photochem Photobiol 51: 621–627

    PubMed  Google Scholar 

  • Hunter CN, van Grondelle R and Olsen JD (1989) Photosynthetic antenna proteins: 100 ps before photochemistry starts. Trends Biol Sci 14: 72–76

    Article  Google Scholar 

  • Jensen A, Aasmundrud O and Eimhjellen KE (1964) Chlorophylls of photosynthetic bacteria. Biochim Biophys Acta 88: 466–479

    PubMed  Google Scholar 

  • Jirsakova V, Agalidis I and Reiss-Husson F (1992) Characterization of the core light-harvesting complex B875 of Rhodocyclus gelatinosus and its B820 derivative. In: Murata N (ed) Research in Photosynthesis, Vol I, pp 33–36. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Kerfeld C, Thornber P and Yeates T (1992) Biochemical and structural characterization of the photosynthetic apparatus of a purple-sulfur bacterium: Chromatium purpuratum. In: Murata N (ed) Research in Photosynthesis, Vol I, pp 49–52. Kluwer Academic Publishers. Dordrecht, The Netherlands

    Google Scholar 

  • Koyama Y (1991) Structures and functions of carotenoids in photosynthetic systems. J Photochem Photbiol B:Biol 9: 265–280

    Article  Google Scholar 

  • Loach PA, Parkes-Loach PS, Davis CM, and Heller BA (1994) Probing protein structural requirements for formation of the core light-harvesting complex of photosynthetic bacteria using hybrid reconstitution methodology. Photosynth Res 40: 231–245 (this issue)

    Google Scholar 

  • Meckensteck RU, Brunisholz RA and Zuber H (1992) The light-harvesting core-complex and the B820-subunit from Rhodopseudomonas marina. Part I. Purification and characterization. FEBS Lett 311: 128–134

    Article  PubMed  Google Scholar 

  • Miller JF, Hinchigeri SB, Parkes-Loach PS, Callahan PM, Sprinkle JR, Riccobono JR and Loach PA (1987) Isolation and characterization of a subunit form of the light-harvesting complex of Rhodospirillum rubrum. Biochemistry 26: 5055–5062

    PubMed  Google Scholar 

  • Miller KR (1982) Three-dimensional structure of a photosynthetic membrane. Nature (London) 300: 53–55

    Google Scholar 

  • Nunn RS, Artymiuk PJ, Baker PJ, Rice DW and Hunter CN (1992) Purification and crystallization of the light harvesting LH1 complex from Rhodobacter sphaeroides. J Mol Biol 228: 1259–1262

    PubMed  Google Scholar 

  • Parkes-Loach PS, Sprinkle JR, and Loach PA (1988) Reconstitution of the B873 light-harvesting complex of Rhodospirillum rubrum from the separately-isolated α- and β-polypeptides and bacteriochlorophyll a. Biochemistry 27: 2718–2727

    PubMed  Google Scholar 

  • Parkes-Loach PS, Michalski TJ, Bass WJ, Smith U and Loach PA (1990) Probing the bacteriochlorophyll binding site by reconstitution of the light-harvesting complex of Rhodospirillum rubrum with bacteriochlorophyll a analogues. Biochemistry 29: 2951–2960

    PubMed  Google Scholar 

  • Pearistein RM (1987) Structure and exciton effects in photosynthesis. In: Amesz J (ed) Photosynthesis, pp 299–317. Elsevier, Amsterdam

    Google Scholar 

  • Pearistein RM (1988) Interpretation of optical spectra of bacteriochlorophyll antenna complexes. In: Scheer H and Schneider S (eds) Photosynthetic Light-Harvesting Systems, pp 555–566. Walter de Gruyter & Co. New York

    Google Scholar 

  • Peters J, Welte W and Drews G (1984) Topographical relationships of polypeptides in the photosynthetic membrane of Rhodopseudomonas viridis investigated by reversible chemical crosslinking. FEBS Lett 171: 267–270

    Article  Google Scholar 

  • Picorel R, Bélanger G and Gingras G (1983) Antenna holochrome B880 of Rhodospirillum rubrum SI. Pigment, phospholipid, and polypeptide composition. Biochemistry 22: 2491–2497

    Google Scholar 

  • Picorel R, L'Ecuyer A, Potier M and Gingras G (1986) Structure of the B880 holochrome of Rhodospirillum rubrum as studied by the radiation inactivation method. J Biol Chem 261: 3020–3024

    PubMed  Google Scholar 

  • Robert B and Lutz M (1990) Structure of the antenna complexes from purple bacteria as seen from resonance Raman spectroscopy. FEMS Symp 53 (Mol Biol Membr-Bound Complexes Phototrophic Bact): 193–198

    Google Scholar 

  • Robert B, Vermeglio A, Steiner R, Scheer H, and Lutz M (1988) BChl a/b in antenna complexes of purple bacteria. In: Scheer H and Schneider S (eds) Photosynthetic Light-Harvesting Systems, pp 355–363. Walter de Gruyter & Co, New York

    Google Scholar 

  • Sauer K (1978) Photosynthetic membranes. Acc Chem Res 11: 257–264

    Google Scholar 

  • Scherz A and Parson WW (1986) Interactions of the bacteriochlorophylls in antenna bacteriochlorophyll-protein complexes of photosynthetic bacteria. Photosynth Res 9: 21–32

    Google Scholar 

  • Shuvalov VA and Asadov AA (1979) Arrangement and interaction of pigment molecules in reaction centers of Rhodospeudomonas viridis. Photodichroism and circular dichroism of reaction centers at 100 K. Biochim Biophys Acta 545: 296–308

    PubMed  Google Scholar 

  • Stark W, Kühlbrandt W, Wildhaber I, Wehrli E and Mühlethaler K (1984) The structure of the photoreceptor unit of Rhodopseudomonas viridis. EMBO J 3: 777–783

    Google Scholar 

  • Stark W, Jay F and Mühlethaler K (1986) Localisation of reaction centre and light harvesting complexes in the photosynthetie unit of Rhodopseudomonas viridis. Arch Microbiol 146: 130–133

    Google Scholar 

  • Steiner R, Cmiel E and Scheer H (1983) Chemistry of bacteriochlorophyll b: Identification of some (photo)oxidation products. Z Naturforsch C 38: 748–752

    Google Scholar 

  • Thompson MA and Fajer J (1992) Calculations of bacteriochlorophyll g primary donors in photosynthetic heliobacteria. How to shift the energy of the phototrap by 2000 cm-1. J Phys Chem 96: 2933–2935

    Article  Google Scholar 

  • Thompson MA, Zerner MC and Fajer J (1991) A theoretical examination of the electronic structure and excited states of the bacteriochlorophyll b dimer from Rhodopseudomonas viridis. J Phys Chem 95: 5693–5700

    Article  Google Scholar 

  • Thornber JP (1971) The photochemical reaction centre of Rhodopseudomonas viridis. Meth Enzymol 23: 688–691

    Google Scholar 

  • Thornber JP, Trosper TL and Strouse CE (1978) Bacteriochlorophyll in vivo: Relationship of spectral forms to specific membrane components. In: Clayton RK and Sistrom WR (eds) The Photosynthetic Bacteria, pp 133–160. Plenum Press, New York

    Google Scholar 

  • Thornber JP, Cogdell RJ, Pierson BK, and Seftor REB (1983) Pigment-protein complexes of purple photosynthetic bacteria: An overview. J Cell. Biochem 23: 159–169

    PubMed  Google Scholar 

  • Van Grondelle R (1985) Excitation energy transfer, trapping and annihilation in photosynthetic systems. Biochim Biophys Acta 811: 147–195

    Google Scholar 

  • Van Mourik F, Visschers RW, Chang MC, Cogdell RJ, Sundström V and van Grondelle R (1990) Spectroscopic properties of pigment-protein complexes from photosynthetic purple bacteria in relation to their structure and function. FEMS Symp S 3 (Mol Biol Membr-Bound Complexes Phototrophic Bact): 345–364

    Google Scholar 

  • Van Mourik F, Corten EPM, van Stokkum IHM, Visschers RW, Loach PA, Kraayenhof R and van Grondelle R (1992) Self assembly of the LH-1 antenna of Rhodospirillum rubrum: A time-resolved study of the aggregation state of the B820 subunit form. In: Murata N (ed) Research in Photosynthesis Vol I, pp 101–104. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Visschers RW, Chang MC, van Mourik F, Parkes-Loach PS, Heller BA, Loach PA and van Grondelle R (1991) Fluorescence polarization and low-temperature absorption spectroscopy of a subunit form of light-harvesting complex I from purple photosynthetic bacteria. Biochemistry 30: 5734–5742

    PubMed  Google Scholar 

  • Wagner-Huber R, Brunisholz RA, Bissig I, Frank G, Suter F and Zuber H (1992) The primary structure of the antenna polypeptides of Ectothiorhodospira halochloris and Ectothiorhodospira halophila. Four core-type antenna polypeptides in E. halochloris and E. halophila. Eur J Biochem 205: 917–925

    PubMed  Google Scholar 

  • Zuber H and Brunisholz RA (1991) Structure and function of antenna polypeptides and chlorophyll-protein complexes: Principles and variability. In: Scheer H (ed) Chlorophylls, pp 627–703 CRC Press, Boca Raton. FL

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, 2153 Sheridan Road, 60208-3500, Evanston, IL, USA

    Pamela S. Parkes-Loach, Sandra M. Jones & Paul A. Loach

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  1. Pamela S. Parkes-Loach
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  2. Sandra M. Jones
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  3. Paul A. Loach
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Parkes-Loach, P.S., Jones, S.M. & Loach, P.A. Probing the structure of the core light-harvesting complex (LH1) of Rhodopseudomonas viridis by dissociation and reconstitution methodology. Photosynth Res 40, 247–261 (1994). https://doi.org/10.1007/BF00034774

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