title: Membrane labeling of coral gastrodermal cells by biotinylation: the proteomic identification of surface proteins involving cnidaria-dinoflagellate endosymbiosis.
abstract: The cellular and molecular-scale processes underlying the stability of coral-Symbiodinium endosymbioses remain unclear despite decades of investigation. As the coral gastroderm is the only tissue layer characterized by this unique symbiotic association, the membranes of these symbiotic gastrodermal cells (SGCs) may play important roles in the initiation and maintenance of the endosymbiosis. In order to elucidate the interactions between the endosymbiotic dinoflagellates and their coral hosts, a thorough characterization of SGC membranes is therefore required. Cell surface proteins of isolated SGCs were biotinylated herein by a cell impermeant agent, biotin-XX sulfosuccinimidyl ester. The in situ distribution of these biotinylated proteins was uncovered by both fluorescence and transmission electron microscopic imaging of proteins bound to Alexa Fluor® 488-conjugated streptavidin. The identity of these proteins was then determined by two-dimensional gel electrophoresis followed by liquid chromatography-tandem mass spectrometry. Nineteen (19) proteins were identified, and they are known to be involved in the molecular chaperone/stress response, cytoskeletal remodeling, and energy metabolism. These results not only reveal the molecular characters of the host SGC membrane, but also provide critical insight into understanding the possible role of host membranes in this ecologically important endosymbiotic association.
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung, Taiwan ; Taiwan Coral Research Center, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan.
authors.forename: Hsing-Hui
authors.initials: HH
authors.lastname: Li
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung, Taiwan.
authors.forename: Zi-Yu
authors.initials: ZY
authors.lastname: Huang
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung, Taiwan.
authors.forename: Shih-Png
authors.initials: SP
authors.lastname: Ye
authors.affiliation: Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
authors.forename: Chi-Yu
authors.initials: CY
authors.lastname: Lu
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
authors.forename: Pai-Chiao
authors.initials: PC
authors.lastname: Cheng
authors.affiliation: Institute of Information Science, Academia Sinica, Taipei, Taiwan.
authors.forename: Shu-Hwa
authors.initials: SH
authors.lastname: Chen
authors.affiliation: Graduate Institute of Marine Biotechnology, National Dong Hwa University, Pingtung, Taiwan ; Taiwan Coral Research Center, National Museum of Marine Biology and Aquarium, Pingtung, Taiwan ; Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, Kaohsiung, Taiwan.
authors.forename: Chii-Shiarng
authors.initials: CS
authors.lastname: Chen
mesh_terms: D000818:Animals; D037421:Anthozoa / Q000469:parasitology / Q000502:physiology*; D019921:Biotinylation; D002462:Cell Membrane / Q000378:metabolism*; D004141:Dinoflagellida / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D013194:Staining and Labeling; D013559:Symbiosis
chemical_list: D008565:Membrane Proteins; D020543:Proteome