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	Characterization of two isoforms of antiliopolysacchride factors (Sp-ALFs) from the mud crab Scylla paramamosain
	Liu, Hai-peng ; Liu HP(刘海鹏) ; Chen, Rong-yuan ; Chen RY(陈荣源) ; Zhang, Qiu-xia ; Zhang QX(张秋霞) ; Wang, Qiao-yang ; Li, Cong-ru ; Peng, Hui ; Peng H(彭会) ; Cai, Ling ; Cai L(蔡灵) ; Zheng, Chao-qun ; Zheng CQ(郑超群) ; Wang, Ke-jian ; Wang KJ(王克坚)
	2012-07
关键词	Antiliopolysacchride factor (ALF) Scylla paramamosain Antimicrobial peptides (AMPs) Antiviral activity
英文摘要	In the previous study of the mud crab (Scylla paramamosain) hemocyte proteins, which interacted with a bacterium, Vibrio parahaemolyticus, a protein known as antilipopolysaccharide factor (Sp-ALF) was isolated in addition to a serine proteinase homolog (Sp-SPH) protein. In the present study, we further reported the characterization of two isoforms of the mud crab ALF - Sp-ALFs genes (designated as Sp-ALF1 and Sp-ALF2, respectively) based on our previous result. The Sp-ALF1 and Sp-ALF2 cDNA contained 1070 bp and 731 bp, respectively, with 123 deduced amino acid residues. Alignment of deduced amino acid sequences showed that Sp-ALFs possessed high identity with other known ALFs from crustaceans and exhibited an overall similarity of 57.7% to those of ALFs compared. Phylogenetic tree analysis revealed a clear group of each species and also suggested that ALFs from Scylla genus and those from Portunus genus were closely related. Tissue distribution analysis in adult crab implied that both Sp-ALF1 and Sp-ALF2 were mainly expressed in hemocytes. The mRNA transcripts were also found in embryo (I, II, III and V), zoea-I and juvenile crab, but were rarely observed in the megalopa stage. To further identify the biological activity of Sp-ALFs, recombinant proteins (rSp-ALFs: designated as rSp-ALF1 and rSp-ALF2, respectively) were obtained by expression in Pichia pastris, and the synthetic peptide fragments (sSp-ALFs: designated as sSp-ALF1 and sSp-ALF2, respectively) including the putative LPS binding loop were also prepared for antimicrobial test. The results indicated that both rSp-ALFs and sSp-ALFs were highly effective against most of the Gram-positive bacteria and Gram-negative bacteria tested. In contrast to cecropin P1, a membrane integrity assay revealed that Sp-ALFs did not affect the Escherichia coil by disruption of membrane integrity. Additionally, the recombinant Sp-ALFs proteins exhibited strong antiviral activity against an important aquaculture pathogen, white spot syndrome virus, in crustaceans. Taken together, these data suggested that Sp-ALFs might play a key role in immune defense against microbial infection in the mud crab S. paramamosain. (C) 2012 Elsevier Ltd. All rights reserved.; The National Natural Science Foundation of China [41006077]; The Scientific Research Starting Foundation for Returned Overseas Chinese Scholars; Ministry of Education, China; Science Foundation of the Fujian Province, China [2010N0030, 2010NZ0002-3, JA10002]; Innovative Research Team in University of Ministry of Education of China [IRT0941]
语种	英语
出版者	ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
内容类型	期刊论文
源URL	[http://dx.doi.org/10.1016/j.fsi.2012.03.014]
专题	海洋环境－已发表论文
推荐引用方式 GB/T 7714	Liu, Hai-peng,Liu HP,Chen, Rong-yuan,et al. Characterization of two isoforms of antiliopolysacchride factors (Sp-ALFs) from the mud crab Scylla paramamosain[J],2012.
APA	Liu, Hai-peng.,刘海鹏.,Chen, Rong-yuan.,陈荣源.,Zhang, Qiu-xia.,...&王克坚.(2012).Characterization of two isoforms of antiliopolysacchride factors (Sp-ALFs) from the mud crab Scylla paramamosain..
MLA	Liu, Hai-peng,et al."Characterization of two isoforms of antiliopolysacchride factors (Sp-ALFs) from the mud crab Scylla paramamosain".(2012).

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