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Isolation and characterization of outer and inner envelope membranes of cyanelles from a glaucocystophyte, Cyanophora paradoxa

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Abstract

Envelope membranes were isolated by sucrose density gradient floatation centrifugation from the homogenate of cyanelles prepared from Cyanophora paradoxa. Two yellow bands were separated after 40 h of centrifugation. The buoyant density of one of the two fractions (fraction Y2) coincided with that of inner envelope membranes of spinach or plasma membranes of cyanobacteria. The other yellow fraction (fraction Y1) migrated to top of sucrose-gradient even at 0% sucrose. Pigment analysis revealed that the heavy yellow fraction was rich in zeaxanthin while the light fraction was rich in β-carotene, and the both fractions contained practically no chlorophylls. Another yellow fraction (fraction Y3) was isolated from the phycobiliprotein fraction, which was the position where the sample was placed for gradient centrifugation. Its buoyant density and absorption spectra were similar to outer membranes of cyanobacteria. We have assigned fractions Y2 and Y3 as inner and outer envelope membrane fractions of cyanelles, respectively. Protein compositions were rather different between the two envelope membranes indicating little cross-contamination among the fractions.

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Abbreviations

DDM:

n-Dodecyl-β-d-maltoside

EDTA:

Ethylenediaminetetraacetic acid

EGTA:

O, O′-Bis(2-aminoethyl)ethyleneglycol-N,N,N′,N′-tetraacetic acid

HPLC:

High performance liquid chromatography

sarkosyl:

Lauroyl sarcosine

SDS-PAGE:

Sodium dodecylsulfate polyacrylamide gel electrophoresis

References

  • Block MA, Dorne AJ, Joyard J, Douce R (1983) Preparation and characterization of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts. II. Biochemical characterization. J Biol Chem 258:13281–13286

    CAS  PubMed  Google Scholar 

  • Cline K, Andrew J, Mersey B, Newcomb E, Keegstra K (1981) Separation and characterization of of inner and outer envelope membranes of pea chloroplasts. Proc Natl Acad Sci USA 78:3595–3599

    Article  CAS  PubMed  Google Scholar 

  • Filip C, Fletcher G, Wulff JL, Earhat CR (1973) Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol 115:717–722

    CAS  PubMed  Google Scholar 

  • Flügge UI, Fischer K, Gross A, Sebald W, Lottspeich F, Eckderskorn C (1989) The triose phosphate-3-phosphoglycerate-phosphate translocator from spinach chloroplast: nucleotide sequence of a full-length cDNA clone and import of the in vitro synthesized precursor protein into chloroplast. EMBO J 8:39–46

    PubMed  Google Scholar 

  • Jakowitsch J, Neumann-Spallart C, Ma Y, Steiner J, Schenk HE, Bohnert HJ, Löffelhardt W (1996) In vitro import of pre-ferredoxin-NADP+-oxidoreductase from Cyanophora paradoxa into cyanelles and into pea chloroplasts. FEBS Lett 381:153–155

    Article  CAS  PubMed  Google Scholar 

  • Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S, Kimura T, Hosouchi T, Matsuno A, Muraki A, Nakazaki N, Naruo K, Okumura S, Shimpo S, Takeuchi C, Wada T, Watanabe A, Yamada M, Yasuda M, Tabata S (1996) Sequence analysis of the genome of the unicellular cyanobacterium Synechocyctis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res 3:109–136

    Article  CAS  PubMed  Google Scholar 

  • Kaneko T, Nakamura Y, Wolk CP, Kuritz T, Sasamoto S, Watanabe A, Iriguchi A, Ishikawa A, Kawashima K, Kimura T, Kishida Y, Kohara M, Matsumoto M, Matsuno A, Muraki A, Nakazaki N, Shimpo S, Sugimoto M, Takazawa M, Yamada M, Yasuda M, Tabata S (2001) Complete genomic sequence of the filamentous nitrogen-fixing cyanobacterium Anabaena sp. strain PCC 7120 DNA Res 8:205–213

    Article  CAS  PubMed  Google Scholar 

  • Kashino Y, Fujimoto K, Akamatsu A, Koike H, Satoh K, Kudoh S (1998) Photosynthetic pigment composition of ice algal and phytoplankton assemblages in early spring in Saroma-ko lagoon, Hokkaido, Japan. Proc NIPR Symp Polar Biol 11:22–32

    Google Scholar 

  • Kashino Y, Koike H, Satoh K (2001) An improved sodium dodecyl sulfate-polyacrylamide gel electrophoresis system to the analysis of membrane protein complexes. Electrophoresis 22:1004–1007

    Article  CAS  PubMed  Google Scholar 

  • Koike H, Ikeuchi M, Hiyama T, Inoue Y (1989) Identification of photosystem I components from the cyanobacterium, Synechococcus vulcanus by N-terminal sequencing. FEBS Lett 253:257–263

    Article  CAS  PubMed  Google Scholar 

  • Koike H, Yoshio M, Kashino Y., Satoh K. (1998) Polypeptide composition of envelopes of spinach chloroplasts: Two major proteins occupy 90% of outer envelope membranes. Plant Cell Physiol 39:526–532

    CAS  PubMed  Google Scholar 

  • Koike H, Shibata M, Yasutomi K, Kashino Y, Satoh K (2000) Identification of photosystem I components from a Glaucocystophyte, Cyanophora paradoxa, by N-terminal sequencing: the PsaD protein is homologous to that of higher plants. Photosynth Res 65:207–217

    Article  CAS  PubMed  Google Scholar 

  • Löffelhardt W, Bohnert HJ (1994) Molecular biology of cyanelles. In: Bryant DA (ed) The molecular biology of cyanobacteria. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 65–89

    Google Scholar 

  • Löffelhardt W, Bohnert HJ, Bryant DA (1997) The cyanelles of Cyanophora paradoxa. Critical Rev Plant Sci 16:393–413

    Article  Google Scholar 

  • Martin W, Bettina S, Goremykin V, Hansmann S, Hasegawa M, Kowallik KV (1998) Gene transfer to the nucleus and the evolution of chloroplasts. Nature 393:162–165

    Article  CAS  PubMed  Google Scholar 

  • Matsuzaki M, Misumi O, Shin-I T, Maruyama S, Takahara M, Miyagishima SY, Mori T, Nishida K, Yagisawa F, Nishida K, Yoshida Y, Nishimura Y, Nakao S, Kobayashi T, Momoyama Y, Higashiyama T, Minoda A, Sano M, Nomoto H, Oishi K, Hayashi H, Ohta F, Nishizaka S, Haga S, Miura S, Morishita T, Kabeya Y, Terasawa K, Suzuki Y, Ishii Y, Asakawa S, Takano H, Ohta N, Kuroiwa H, Tanaka K, Shimizu N, Sugano S, Sato N, Nozaki H, Ogasawara N, Kohara Y, Kuroiwa T. (2004) Genome sequence of the ultrasmall unicellular red alga Cyanidioschyzon merolae 10D. Nature 428:653–657

    Article  CAS  PubMed  Google Scholar 

  • McFadden GI (1999) Endosymbiosis and evolution of the plant cell. Curr Opinion Plant Biol 2:513–519

    Article  CAS  Google Scholar 

  • Murata N, Sato N, Omata T, Kuwabara T (1981) Separation and characterization of thylakoid and cell envelope of the blue-green alga (cyanobacterium) Anasystis nidulans. Plant Cell Physiol 22:855–866

    CAS  Google Scholar 

  • Nozaki H, Matsuzaki M, Takahara M, Misumi O, Kuroiwa H., Hasegawa M, Shin-I T., Kohara Y, Ogasawara N, Kuroiwa T (2003) The phylogenetic position of red algae revealed by multiple nuclear genes from mitochondria-containing eukaryotes and an alternative hypothesis on the origin of plastids. J Mol Evol 56:485–497

    Article  CAS  PubMed  Google Scholar 

  • Omata T, Murata N (1983) Isolation and characterization of the cytoplasmic membranes from the blue-green alga (cyanobacterium) Anacystis nidulans. Plant Cell Physiol 24:1101–1112

    CAS  Google Scholar 

  • Omata T, Murata N (1984) Isolation and characterization of three types of membranes from the cyanobacterium (blue-green alga) Synechocystis sp.PCC6714. Arch Microbiol 139:113–116

    Article  CAS  Google Scholar 

  • Omata T, Murata N (1985) Separation and partial characterization of membranes from Prochloron sp. Plant Cell Physiol 26:579–584

    CAS  Google Scholar 

  • Pfanzgle B, Löffelhardt W (1999) In vitro synthesis of peptideglycan precursors modified with N-acetylputresine by Cyanophora paradoxa cyanelles envelope membranes. J Bacteriol 181:2643–2647

    Google Scholar 

  • Reumann S, Davila-Aponte J, Keegstra K (1999) The evolutionary origin of the protein-translocating channel chloroplastic envelope membranes: identification of a cyanobacterial homolog. Proc Natl Acad Sci USA 96:784–789

    Article  CAS  PubMed  Google Scholar 

  • Schlichting R, Zimmer W, Bothe H. (1990) Exchange of metabolites in Cyanophora paradoxa and its cyanelles. Bot Acta 103:392–398

    Google Scholar 

  • Shibata M, Kashino Y, Satoh K, Koike H (2001) Isolation and characterization of Oxygen-evolving Thylakoid membranes and photosystem II particles from a Glaucocystophyte, Cyanophora paradoxa. Plant Cell Physiol 42:733–741

    Article  CAS  PubMed  Google Scholar 

  • Steiner JM, Löffelhardt W (2002) Protein import into cyanelles. Trends Plant Sci 7:72–77

    Article  CAS  PubMed  Google Scholar 

  • Stirewalt VL, Michalowski CB, Löffelhardt W, Bohnert HJ, Bryant DA (1995) Nucleotide sequence of the cyanelle genome from Cyanophora paradoxa. Plant Mol Biol Reptr 13:327–332

    Article  CAS  Google Scholar 

  • Wilhelm SW, Trick CG (1995) Alterations in cell pigmentation, protein expression, and photosynthetic capacity of the cyanobacterium Oscillatoria tenuis grown under low iron conditions. Can J Microbiol 41:145–151

    Article  CAS  Google Scholar 

  • Yusa F, Kashino Y, Satoh K and Koike H (2001) Isolation and characterization of envelope membranes from cyanelles of Cyanophora paradoxa. In: PS2001 Proceedings of 12th international congress of photosynthesis, S23-012, CSIRO Publishing, Melbourne (CD-ROM)

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Acknowledgments

The authors wish to thank Dr. E. Yokota, University of Hyogo, for preparation and observation of electron microscopy analysis. This work was supported in part by a Grant-in-aid for the scientific research from The Ministry of Education, Science, Sports and Culture Japan to HK (No. 11640655) and by a grant of 21st Century of Excellence to KS.

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Authors and Affiliations

  1. Department of Life Science, Graduate School of Life Science, University of Hyogo, Ako, Hyogo, 678-1297, Japan

    Hiroyuki Koike, Yohei Ikeda, Fumie Yusa, Yasuhiro Kashino & Kazuhiko Satoh

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  1. Hiroyuki Koike
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  2. Yohei Ikeda
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  3. Fumie Yusa
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  4. Yasuhiro Kashino
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  5. Kazuhiko Satoh
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Correspondence to Hiroyuki Koike.

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H. Koike and Y. Ikeda contributed equally.

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Koike, H., Ikeda, Y., Yusa, F. et al. Isolation and characterization of outer and inner envelope membranes of cyanelles from a glaucocystophyte, Cyanophora paradoxa . Photosynth Res 93, 45–53 (2007). https://doi.org/10.1007/s11120-007-9156-2

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  • DOI: https://doi.org/10.1007/s11120-007-9156-2

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