Four distinct trimeric forms of light-harvesting complex II isolated from the green alga Chlamydomonas reinhardtii

  • Keisuke KawakamiEmail author
  • Ryutaro Tokutsu
  • Eunchul Kim
  • Jun Minagawa
Original article


Light-harvesting complex II (LHCII) absorbs light energy and transfers it primarily to photosystem II in green algae and land plants. Although the trimeric structure of LHCII is conserved between the two lineages, its subunit composition and function are believed to differ significantly. In this study, we purified four LHCII trimers from the green alga Chlamydomonas reinhardtii and analyzed their biochemical properties. We used several preparation methods to obtain four distinct fractions (fractions 1–4), each of which contained an LHCII trimer with different contents of Type I, III, and IV proteins. The pigment compositions of the LHCIIs in the four fractions were similar. The absorption and fluorescence spectra were also similar, although the peak positions differed slightly. These results indicate that this green alga contains four types of LHCII trimer with different biochemical and spectroscopic features. Based on these findings, we discuss the function and structural organization of green algal LHCII antennae.


Chlamydomonas Light-harvesting complex II trimer Photosystem II Subunit composition 



Coomassie brilliant blue




Column volume






Light-harvesting complex II


2-(N-morpholino) ethanesulfonic acid


Photosystem II


Photosystem I


Sucrose density gradient


Sodium dodecyl sulfate–polyacrylamide gel electrophoresis


Ultra-performance liquid chromatography



We thank Ms. Rie Uno (Osaka City University), Mr. Daisuke Namba (Osaka City University), and Ms. Chiyo Noda (National Institute for Basic Biology) for their assistance with purification and analysis. We are also grateful to Prof. Nobuo Kamiya (Osaka City University) and Associate Prof. Ikuko Miyahara (Osaka City University) for providing laboratory access. This work was funded by the Joint Usage/Research Program of the Artificial Photosynthesis, Osaka City University. K.K. and J.M. are grateful for the continued support of JST CREST, Japan (Grant No. JPMJCR13M4).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Caffarri S, Croce R, Cattivelli L, Bassi R (2004) A look within LHCII: differential analysis of the Lhcb1-3 complexes building the major trimeric antenna complex of higher-plant photosynthesis. Biochemistry 43:9467–9476CrossRefGoogle Scholar
  2. Drop B, Webber-Birungi M, Yadav SKN, Filipowicz-Szymanska A, Fusetti F, Boekema EJ, Croce R (2014) Light-harvesting complex II (LHCII) and its supramolecular organization in Chlamydomonas reinhardtii. Biochim Biophys Acta 1837:63–72CrossRefGoogle Scholar
  3. Elrad D, Niyogi KK, Grossman AR (2002) A major light-harvesting polypeptide of photosystem II functions in thermal dissipation. Plant Cell 14:1801–1816CrossRefGoogle Scholar
  4. Ferrante P, Ballottari M, Bonente G, Giuliano G, Bassi R (2012) LHCBM1 and LHCBM2/7 polypeptides, components of major LHCII complex, have distinct functional roles in photosynthetic antenna system of Chlamydomonas reinhardtii. J Biol Chem 287:16276–16288CrossRefGoogle Scholar
  5. García-Cerdán JG, Kovács L, Tõth T et al (2011) The PsbW protein stabilizes the supramolecular organization of photosystem II in higher plants. Plant J 65:368–381CrossRefGoogle Scholar
  6. Gorman DS, Levine RP (1965) Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Natl Acad Sci USA 54:1665–1669CrossRefGoogle Scholar
  7. Grewe S, Ballottari M, Alcocer M et al (2014) Light-harvesting complex protein LHCBM9 is critical for photosystem II activity and hydrogen production in Chlamydomonas reinhardtii. Plant Cell 26:1598–1611CrossRefGoogle Scholar
  8. Gulis G, Narasimhulu KV, Redding KE (2008) Purification of His(6)-tagged photosystem I from Chlamydomonas reinhardtii. Photosynth Res 96:51–60CrossRefGoogle Scholar
  9. Ikeuchi M, Inoue Y (1988) A new 4.8-kDa polypeptide intrinsic to the PS II reaction center, as revealed by modified SDS-PAGE with improved resolution of low-molecular-weight proteins. Plant Cell Physiol 29:1233–1239Google Scholar
  10. Jackowski G, Kacprzak K, Jansson S (2001) Identification of Lhcb1/Lhcb2/Lhcb3 heterotrimers of the main light-harvesting chlorophyll a/bprotein complex II of Photosystem II (LHCII). Biochim Biophys Acta 1504:340–345CrossRefGoogle Scholar
  11. Janik E, Bednarska J, Sowinski K, Luchowski R, Zubik M, Grudzinski W, Gruszecki WI (2017) Light-induced formation of dimeric LHCII. Photosynth Res 132:265–276CrossRefGoogle Scholar
  12. Jansson S (1999) A guide to the Lhc genes and their relatives in Arabidopsis. Trends Plant Sci 4:236–240CrossRefGoogle Scholar
  13. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685CrossRefGoogle Scholar
  14. Lamb JJ, Røkke G, Hohmann-Marriott MF (2018) Chlorophyll fluorescence emission spectroscopy of oxygenic organisms at 77 K. Photosynthetica 56:105–124CrossRefGoogle Scholar
  15. Liu Z, Yan H, Wang K, Kuang T, Zhang J, Gui L, An X, Chang W (2004) Crystal structure of spinach major light-harvesting complex at 2.72 Å resolution. Nature 428:287–292CrossRefGoogle Scholar
  16. Minagawa J, Takahashi Y (2004) Structure, function and assembly of Photosystem II and its light-harvesting proteins. Photosynth Res 82:241–263CrossRefGoogle Scholar
  17. Natali A, Croce R (2015) Characterization of the major light-harvesting complexes (LHCBM) of the green alga Chlamydomonas reinhardtii. PLoS ONE 10:1–18CrossRefGoogle Scholar
  18. Nguyen AV, Thomas-Hall SR, Malnoë A et al (2008) Transcriptome for photobiological hydrogen production induced by sulfur deprivation in the green alga Chlamydomonas reinhardtii. Eukaryot Cell 7:1965–1979CrossRefGoogle Scholar
  19. Standfuss J, Van Scheltinga ACT, Lamborghini M, Kühlbrandt W (2005) Mechanisms of photoprotection and nonphotochemical quenching in pea light-harvesting complex at 2.5 Å resolution. EMBO J 24:919–928CrossRefGoogle Scholar
  20. Su X, Ma J, Wei X, Cao P, Zhu D, Chang W, Liu Z, Zhang X, Mei L (2017) Structure and assembly mechanism of plant C2S2M2-type PSII-LHCII supercomplex. Science 357:815–820CrossRefGoogle Scholar
  21. Takahashi H, Iwai M, Takahashi Y, Minagawa J (2006) Identification of the mobile light-harvesting complex II polypeptides for state transitions in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 103:477–482CrossRefGoogle Scholar
  22. Takahashi H, Okamuro A, Minagawa J, Takahashi Y (2014) Biochemical characterization of photosystem I-associated light-harvesting complexes I and II isolated from state 2 cells of Chlamydomonas reinhardtii. Plant Cell Physiol 55:1437–1449CrossRefGoogle Scholar
  23. Tokutsu R, Minagawa J (2013) Energy-dissipative supercomplex of photosystem II associated with LHCSR3 in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 110:10016–10021CrossRefGoogle Scholar
  24. Tokutsu R, Kato N, Bui KH, Ishikawa T, Minagawa J (2012) Revisiting the supramolecular organization of photosystem II in Chlamydomonas reinhardtii. J Biol Chem 287:31574–31581CrossRefGoogle Scholar
  25. Wei X, Su X, Cao P, Liu X, Chang W, Li M, Zhang X, Liu Z (2016) Structure of spinach photosystem II-LHCII supercomplex at 3.2 Å resolution. Nature 534:69–74CrossRefGoogle Scholar
  26. Wittig I, Braun HP, Schägger H (2006) Blue native PAGE. Nat Protoc 1:418–428CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.The OCU Advanced Research Institute for Natural Science & Technology (OCARINA)Osaka City UniversityOsaka CityJapan
  2. 2.Division of Environmental PhotobiologyNational Institute for Basic BiologyMyodaijiJapan

Personalised recommendations