Cloning and characterization of two different ficolins from the giant freshwater prawn Macrobrachium rosenbergii.
Ficolins, a kind of lectin containing collagen-like and fibrinogen-related domains (FReDs, also known as FBG or FREP), are involved in the first line of host defense against pathogens. In this study, two ficolins, namely, MrFico1 and MrFico2, from the giant freshwater prawn Macrobrachium rosenbergii were identified. In contrast to other ficolins, these two ficolins have no collagen-like domain, but such ficolins contain a coiled region and a FReD domain. Phylogenetic analysis showed that MrFico1 and MrFico2, together with two ficolin-like proteins from Pacifastacus leniusculus, belonged to one group. Quantitative RT-PCR (qRT-PCR) showed that both MrFico1 and MrFico2 were expressed in hepatopancreas, stomach and intestine, with the highest expression in stomach for MrFico1, compared to the highest expression in hepatopancreas for MrFico2. qRT-PCR analysis also showed that MrFico1 was obviously upregulated upon Vibrio anguillarium challenge, while MrFico2 was upregulated after challenged by V. anguillarium or white spot syndrome virus. Bacterium-binding experiment showed that MrFico1 and MrFico2 could bind to different microbes, and sugar-binding assay revealed that these two ficolins could also bind to lipopolysaccharide and peptidoglycan, the glycoconjugates of bacteria surface. Moreover, these two ficolins could agglutinate bacteria in a calcium-dependent manner, and the results of bacteria clearance experiment showed that both ficolins could facilitate the clearance of injected bacteria in the prawn. Our results suggested that MrFico1 and MrFico2 may function as pattern-recognition receptors in the immune system of M. rosenbergii.
School of Life Science, Henan Normal University, Xinxiang, Henan 453007, China.
Xiao-Wen
XW
Zhang
The Key Laboratory of Plant Cell Engineering and Germplasm Innovation of Ministry of Education/Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong 250100, China.
Xian-Wei
XW
Wang
Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
Ying
Y
Huang
Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
Kai-Min
KM
Hui
Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
Yan-Ru
YR
Shi
Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China. Electronic address: njnuwang@263.net.
Wen
W
Wang
Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China. Electronic address: renqian0402@126.com.
Qian
Q
Ren
D000371:Agglutination; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004266:DNA Virus Infections / Q000276:immunology*; D043143:Hepatopancreas / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism*; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000276:immunology*; D010457:Peptidoglycan / Q000378:metabolism; D010802:Phylogeny; D011485:Protein Binding; D017434:Protein Structure, Tertiary / Q000235:genetics; D051192:Receptors, Pattern Recognition / Q000378:metabolism; D059467:Transcriptome; D015854:Up-Regulation; D014733:Vibrio / Q000276:immunology*; D014735:Vibrio Infections / Q000276:immunology*; D046848:White spot syndrome virus 1 / Q000276:immunology*; D000097604:Ficolins
D037102:Lectins; D008070:Lipopolysaccharides; D010457:Peptidoglycan; D051192:Receptors, Pattern Recognition; D000097604:Ficolins
Bacteria binding; Bacteria clearance; Ficolin; Innate immunity