Molecular and functional characterization of ferritin in abalone Haliotis diversicolor supertexta
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Molecular and functional characterization of ferritin in abalone Haliotis diversicolor supertexta XIE Jiasong1† , CAO Xiaohua1† , WU Liuji1 , LUO Ming1 , ZHU Zewen1 , HUANG Yanqing1 , WU Xinzhong1∗ 1
Laboratory of Marine Life Science and Technology, College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
Received 25 January 2011; accepted 28 November 2011 ©The Chinese Society of Oceanography and Springer-Verlag Berlin Heidelberg 2012
Abstract Ferritin is an iron storage protein that plays a key role in the processes of physiology and pathology. In the present study, the authors reported the ferritin gene from abalone Haliotis diversicolor supertexta, which we named hds-ferritin. The full-length of hds-ferritin cDNA consisted of 879 bp with an ORF encoding a 171 amino acids. Amino acid sequence analysis revealed that hds-ferritin shared highly homology with other species. Real time PCR and western blot analysis showed that hds-ferritin was distributed ubiquitously in abalone tissues and had the highest expression level in digestive glands, but its transcripts are not modified remarkably by the stimulation with LPS. The recombinant protein was successfully expressed in Escherichia coli BL21 (DE3), and the titre of anti-ferritin antibody was about 1:14 000. The effects of ROS and RNS on ferritin were analyzed in the present study. The results showed that H2 O2 played an important role in decreasing hds-ferritin, however NO cation appeared to have a protecting effect on H2 O2 -medied reduction of hds-ferritin. Key words: Haliotis diversicolor supertexta, ferritin, respiratory burst, oxidative damage, immune response
ferritins (Zoysa and Lee, 2007). It has become evident that ferritin can convert Fe(II) to Fe(III) and its synthesis is normally regulated not only by the intracellular iron concentration, but also by cytokines, oxidants, oncogenes, growth factor and so on (Harrison and Arosio, 1996; Torti and Torti, 2002). More recently, reports show that serum ferritin can regulate vascular remodelling and angiogenesis (Coffman et al., 2009) and the ferritin Fe2 site at the diiron catalytic center can control the reaction with O2 in the rapid mineralization pathway (Tosha et al., 2008). Abalone is an important mollusc species for commercial production in China and in the world, however, the abalone diseases have caused complications for the global abalone industry (Gardner et al., 1995; Moore et al., 2000; Wu, 2003). Therefore, it is important to focus mainly on improving non-specific immune functions of marine invertebrates. As it is well known, respiratory burst is an important physiological function, especially in the non-specific immune system
1 Introduction As a highly conserved iron storage protein, ferritin was found in all species from animals to plants and bacteria. Structurally a ferritin molecule consists of a hollow shell which is composed of 24 subunits and able to store up to 4500 Fe (III) atoms as an inorganic complex (Theil, 1987; Harrison and Arosio, 1996). In vertebrates, 24 subunits assem
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