口虾蛄主要过敏原原肌球蛋白的免疫活性

CORC > 集美大学

	口虾蛄主要过敏原原肌球蛋白的免疫活性
	蔡秋凤 ; 王锡昌 ; 刘光明 ; 罗志湖 ; 曹敏杰
刊名	http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=sckx201003014&dbcode=CJFQ&dbname=CJFQ2010
	2012-06-05 ; 2012-06-05
关键词	口虾蛄原肌球蛋白纯化免疫交叉反应定量 mantis shrimp tropomyosin purification cross-reactivity quantification S917.4
其他题名	Studies on the immunoreactivity of the major allergen tropomyosin in mantis shrimp (Squilla oratoria)
中文摘要	原肌球蛋白(TM)是甲壳类动物的主要过敏原,其氨基酸序列具有很高的保守性。本研究以口虾蛄为研究对象,旨在探明TM是否是其主要过敏原并对其相关性质进行研究。利用过敏患者血清通过免疫印迹法确定口虾蛄的主要过敏原分子量约为36ku。通过制备丙酮粉、等电点沉淀、硫酸铵盐析及加热等方法纯化了该主要过敏原。采用抗中华绒螯蟹TM多克隆抗体进行免疫学性质分析表明该蛋白为TM。抑制性免疫印迹和抑制性ELISA实验表明口虾蛄TM与凡纳滨对虾和锯缘青蟹等甲壳类动物的TM具有较强的免疫交叉反应,但与贝类动物杂色蛤的免疫交叉性较低。通过ELISA方法对甲壳类动物的TM进行定量测定,结果表明,口虾蛄肌肉中TM的含量约只有凡纳滨对虾TM含量的1/45,但仍具有一定的致敏性。; Crustacean is one of the eight kinds of allergen sources in coastal areas,which causes the IgE-mediated hypersensitive reactions with clinical manifestations including urticaria,angioedema,asthma,and even fatal anaphylaxis. Tropomyosin (TM) is the major allergen of decapod crustaceans with highly conserved amino acid sequences. The purpose of this study is to confirm whether TM is a major cross-reactive allergen in mantis shrimp (Squilla oratoria) which is taxonomically distinct from decapods and largely consumed as a delicacy in China. Quantification of TM in different species of crustaceans was also conducted. Muscle sample from mantis shrimp was homogenized with phosphate buffer to prepare heated extracts and separated by 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A protein band with molecular mass of about 36 ku was detected by IgE-immunoblotting by all of the sera with crustacean allergy,suggesting it is the major allergen of mantis shrimp. This protein was further purified to homogeneity by acetone powder preparation,isoelectric point precipitation,ammonium sulfate fractionation (40%-60% saturation),and heat treatment. Purified protein was demonstrated to be TM by Western blot using polyclonal antibody against TM from Chinese mitten crab (Eriocheir sinensis). Heated extracts from other crustaceans including mud crab (Scylla serrata),Pacific white shrimp (Penaeus vannamei) and short necked clam (Venerupis variegata) were also prepared,and the cross-reactivity of TM from mantis shrimp with TMs from these crustaceans was confirmed by inhibition immunoblotting using polyclonal antibody against TM and inhibition ELISA using sera with crustacean allergy. Quantification by ELISA using polyclonal antibody against TM revealed that TM content in mantis shrimp muscle is much lower than that in Pacific white shrimp and mud crab muscle,which was about 45 times lower in mantis shrimp muscle than that in Pacific white shrimp muscle. However,its allergenicity seems equivalent to other decapod crustaceans as showed by inhibition ELISA using sera with crustacean allergy. This may be due to the degradation of TM in mantis shrimp by its endogenous serine proteinases and cathepsins which destroyed the IgG-binding epitope but not the IgE-binding epitopes. In conclusion,this study demonstrated that allergenicity of mantis shrimp is almost equivalent to decapods and TM is the major allergen in mantis shrimp with high cross-reactivity to decapod crustaceans.; 【作者单位】上海海洋大学食品学院；集美大学生物工程学院；福建省高校水产科学技术与食品安全重点实验室；【作者英文名】CAI Qiu-feng 1,2,3 ,WANG Xi-chang 1 ,LIU Guang-ming 2,3 ,LUO Zhi-hu2,CAO Min-jie 2,3 (1. College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306,China; 2. School of Biotechnology Engineering,Jimei University,Xiamen 361021,Chi
语种	中文
内容类型	期刊论文
源URL	[http://ir.calis.edu.cn/hdl/235041/18541]
专题	集美大学
推荐引用方式 GB/T 7714	蔡秋凤,王锡昌,刘光明,等. 口虾蛄主要过敏原原肌球蛋白的免疫活性[J]. http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=sckx201003014&dbcode=CJFQ&dbname=CJFQ2010,2012, 2012.
APA	蔡秋凤,王锡昌,刘光明,罗志湖,&曹敏杰.(2012).口虾蛄主要过敏原原肌球蛋白的免疫活性.http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=sckx201003014&dbcode=CJFQ&dbname=CJFQ2010.
MLA	蔡秋凤,et al."口虾蛄主要过敏原原肌球蛋白的免疫活性".http://epub.edu.cnki.net/grid2008/brief/detailj.aspx?filename=sckx201003014&dbcode=CJFQ&dbname=CJFQ2010 (2012).