Suma Yota, Ishizaki Shoichiro, Nagashima Yuji, Lu Ying, Ushio Hideki, Shiomi Kazuo
Department of Food Science and Technology, Tokyo University of Marine Science and Technology, Konan-4, Tokyo 108-8477, Japan.
Comp Biochem Physiol B Biochem Mol Biol. 2007 Jun;147(2):230-6. doi: 10.1016/j.cbpb.2007.01.004. Epub 2007 Jan 24.
Tropomyosin, a myofibrillar protein of 35-38 kDa, represents a major and cross-reactive allergen in decapod crustaceans. This study was initiated to clarify whether decapod-allergic patients also recognize tropomyosins of barnacles, crustaceans phylogenetically remote from decapods, which are locally consumed as a delicacy. On SDS-PAGE, a 37 kDa protein was observed in all the heated extracts prepared from two species of decapods (American lobster Homarus americanus and black tiger prawn Penaeus monodon) and two species of barnacles (acorn barnacle Balanus rostratus and goose barnacle Capitulum mitella). In immunoblotting, the 37 kDa protein was found to react with monoclonal antibodies against American lobster tropomyosin and hence identified as tropomyosin. The patient sera reacted to tropomyosins from both decapods and barnacles and the reactivity was abolished by preincubation with American lobster tropomyosin, demonstrating that barnacle tropomyosins are allergens cross-reactive with decapod tropomyosins. However, the amino acid sequence of acorn barnacle tropomyosin, deduced by cDNA cloning experiments, shares higher sequence identity with abalone tropomyosins than with decapod tropomyosins. In accordance with this, the phylogenetic tree made for tropomyosins from various animals showed that the acorn barnacle tropomyosin is evolutionally classified not into the decapod tropomyosin family but into the molluscan tropomyosin family.
原肌球蛋白是一种分子量为35 - 38 kDa的肌原纤维蛋白,是十足目甲壳类动物中的主要交叉反应性过敏原。开展本研究是为了弄清楚对十足目动物过敏的患者是否也会识别藤壶的原肌球蛋白,藤壶是与十足目动物在系统发育上距离较远的甲壳类动物,在当地被作为美味食用。在SDS - PAGE上,在从两种十足目动物(美洲龙虾美洲螯龙虾和黑虎虾斑节对虾)以及两种藤壶(纹藤壶和龟甲藤壶)制备的所有加热提取物中均观察到一种37 kDa的蛋白质。在免疫印迹中,发现该37 kDa蛋白质与抗美洲龙虾原肌球蛋白的单克隆抗体发生反应,因此被鉴定为原肌球蛋白。患者血清与十足目动物和藤壶的原肌球蛋白均发生反应,并且通过与美洲龙虾原肌球蛋白预孵育可消除这种反应性,这表明藤壶原肌球蛋白是与十足目动物原肌球蛋白交叉反应的过敏原。然而,通过cDNA克隆实验推导的纹藤壶原肌球蛋白的氨基酸序列与鲍鱼原肌球蛋白的序列同一性高于与十足目动物原肌球蛋白的序列同一性。据此,针对各种动物原肌球蛋白构建的系统发育树表明,纹藤壶原肌球蛋白在进化上并非归类于十足目动物原肌球蛋白家族,而是归类于软体动物原肌球蛋白家族。