title: Identification and characterization of the related immune-enhancing proteins in crab Scylla paramamosain stimulated with rhubarb polysaccharides.
abstract: Recently, considerable interest has been focused on immunostimulants to reduce diseases in crab aquaculture. However, information regarding to the related immune-enhancing proteins in crabs is not available yet. In this study, rhubarb polysaccharides were tested for enhancement of the immune activity in crab Scylla paramamosain. Compared with those in the control group, values of, phenoloxidase (PO), alkaline phosphatase (AKP) and alkaline phosphatasein (ACP) activity in the, experimental group were improved significantly 4 d after the treatment. Furthermore, 15 and 17 altered proteins from haemocytes and hepatopancreas, respectively, were found in rhubarb polysaccharide-treated crabs using 2-DE approach. Of these, hemocyanin, chymotrypsin, cryptocyanin, C-type lectin receptor, and ferritin protein were identified by mass spectrometry. In addition, RT-PCR, analysis showed that the mRNA levels of hemocyanin and chymotrypsin increased about 2.4- and 1.4-fold in the experiment group. Moreover, the hemocyanin gene in S. paramamosain (SpHMC) was, cloned and characterized. SpHMC contains one open reading frame of 2022 bp and encodes a polypeptide of 673 amino acids. It is clustered into one branch along with crab hemocyanin in a phylogenetic tree. The mRNA transcripts of SpHMC were detected mainly in the tissues of, hepatopancreas, hemocyte and intestines, and its levels were up-regulated significantly in hemocytes, of S. paramamosain treated with Vibrio parahemolyticus, Beta streptococcus or poly I:C for 6-48 h. Taken together, these studies found 5 related immune-enhancing proteins and a novel heomcyanin homologue with potential pathogen-resistant activities in crab.
authors.affiliation: Department of Biology and Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou, Guangdong 515063, China.
authors.forename: Jingsong
authors.initials: J
authors.lastname: Cao
authors.forename: Zehuan
authors.initials: Z
authors.lastname: Wang
authors.forename: Yueling
authors.initials: Y
authors.lastname: Zhang
authors.forename: Fengliang
authors.initials: F
authors.lastname: Qu
authors.forename: Lingling
authors.initials: L
authors.lastname: Guo
authors.forename: Mingqi
authors.initials: M
authors.lastname: Zhong
authors.forename: Shengkang
authors.initials: S
authors.lastname: Li
authors.forename: Haiying
authors.initials: H
authors.lastname: Zou
authors.forename: Jiehui
authors.initials: J
authors.lastname: Chen
authors.forename: Xiuying
authors.initials: X
authors.lastname: Wang
mesh_terms: D000276:Adjuvants, Immunologic / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003386:Brachyura / Q000378:metabolism*; D002918:Chymotrypsin / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D006433:Hemocyanins / Q000235:genetics / Q000378:metabolism; D006434:Hemocytes / Q000378:metabolism; D043143:Hepatopancreas / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D014442:Monophenol Monooxygenase / Q000378:metabolism; D028321:Plant Preparations / Q000378:metabolism; D011070:Poly I-C / Q000276:immunology; D011134:Polysaccharides / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D012250:Rheum / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013291:Streptococcus / Q000276:immunology; D014733:Vibrio / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D037181:Lectins, C-Type; D028321:Plant Preparations; D011134:Polysaccharides; D012333:RNA, Messenger; D006433:Hemocyanins; D014442:Monophenol Monooxygenase; D000469:Alkaline Phosphatase; D002918:Chymotrypsin; D011070:Poly I-C
keywords: Hemocyanin; Immunostimulant; Pathogen-resistance; Related immune-enhancing proteins; Scylla paramamosain