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SLDP: a Novel Protein Related to Caleosin Is Associated with the Endosymbiotic Symbiodinium Lipid Droplets from Euphyllia glabrescens

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Abstract

Intracellular lipid droplets (LDs) have been proposed to play a key role in the mutualistic endosymbiosis between reef-building corals and the dinoflagellate endosymbiont Symbiodinium spp. This study investigates and identifies LD proteins in Symbiodinium from Euphyllia glabrescens. Discontinuous Percoll gradient centrifugation was used to separate Symbiodinium cells from E. glabrescens tentacles. Furthermore, staining with a fluorescent probe, Nile red, indicated that lipids accumulated in that freshly isolated Symbiodinium cells and lipid analyses further showed polyunsaturated fatty acids (PUFA) was abundant. The stable LDs were purified from endosymbiotic Symbiodinium cells. The structural integrity of the Symbiodinium LDs was maintained via electronegative repulsion and steric hindrance possibly provided by their surface proteins. Protein extracts from the purified LDs revealed a major protein band with a molecular weight of 20 kDa, which was termed Symbiodinium lipid droplet protein (SLDP). Interestingly, immunological cross-recognition analysis revealed that SLDP was detected strongly by the anti-sesame and anti-cycad caleosin antibodies. It was suggested that the stable Symbiodinium LDs were sheltered by this unique structural protein and was suggested that SLDP might be homologous to caleosin to a certain extent.

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Abbreviations

FSW:

Filtered natural seawater

LD:

Lipid droplet

MLDP:

Major lipid droplet protein

PCR:

Polymerase chain reaction

Q-TOF:

Quadrupole time-of-flight

RFLP:

Restriction fragment length polymorphism

SLDP:

Symbiodinium lipid droplet protein

TAG:

Triacylglycerol

PUFA:

Polyunsaturated fatty acid

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Acknowledgments

The work was supported by a grant from the National Science Council, Taiwan, ROC (NSC 102-2313-B-291-001 to PL Jiang).

Author information

Authors and Affiliations

  1. Graduate Institute of Marine Biotechnology, National Dong-Hwa University, Pingtung, 944, Taiwan

    Chii-Shiarng Chen & Pei-Luen Jiang

  2. Taiwan Coral Research Center, National Museum of Marine Biology and Aquarium, Pingtung, 944, Taiwan

    Tung-Yung Fan, Yu-Min Ju, Jing-O Cheng, Chii-Shiarng Chen & Pei-Luen Jiang

  3. Graduate Institute of Biotechnology, National Chung-Hsing University, Taichung, 402, Taiwan

    Buntora Pasaribu, I-Ping Lin & Jason T. C. Tzen

  4. Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, Kaohsiung, 804, Taiwan

    Chii-Shiarng Chen

  5. School of Chinese Medicine, China Medical University, Taichung, 404, Taiwan

    Jason T. C. Tzen

  6. Agricultural Biotechnology Research Center, Academia Sinica, Taipei, 115, Taiwan

    Jason T. C. Tzen

  7. Institute of Plant and Microbial Biology, Academia Sinica, Taipei, 115, Taiwan

    Guang-Yuh Jauh

Authors
  1. Buntora Pasaribu
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  2. I-Ping Lin
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  9. Pei-Luen Jiang
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Correspondence to Chii-Shiarng Chen or Pei-Luen Jiang.

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Fig. S1

Symbiodinium clade identification by Restriction Fragmentation Length Polymorphism (RFLP). The clade of the Symbiodinium cells was identified by RFLP analysis of the n18S-rDNA. The fragments of n18S-rDNA were digested by TaqI and Sau3AI and then resolved by electrophoresis. The RFLP patterns of the freshly isolated and free-living culture Symbiodinium cells were similar to each other. Both were identified as belonging to clade C. (GIF 204 kb)

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Pasaribu, B., Lin, IP., Tzen, J.T.C. et al. SLDP: a Novel Protein Related to Caleosin Is Associated with the Endosymbiotic Symbiodinium Lipid Droplets from Euphyllia glabrescens . Mar Biotechnol 16, 560–571 (2014). https://doi.org/10.1007/s10126-014-9574-z

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