Yang Yang, Zhang Yong-Xia, Liu Meng, Maleki Soheila J, Zhang Ming-Li, Liu Qing-Mei, Cao Min-Jie, Su Wen-Jin, Liu Guang-Ming
College of Food and Biological Engineering, Xiamen Key Laboratory of Marine Functional Food, Fujian Provincial Engineering Technology Research Center of Marine Functional Food, Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Jimei University , Xiamen, Fujian 361021, China.
Agricultural Research Service, Southern Regional Research Center, U. S. Department of Agriculture , New Orleans, Louisiana 70124, United States.
J Agric Food Chem. 2017 Feb 1;65(4):950-963. doi: 10.1021/acs.jafc.6b04587. Epub 2017 Jan 23.
Triosephosphate isomerase (TIM) is a key enzyme in glycolysis and has been identified as an allergen in saltwater products. In this study, TIM with a molecular mass of 28 kDa was purified from the freshwater crayfish (Procambarus clarkii) muscle. A 90-kDa protein that showed IgG/IgE cross-reactivity with TIM was purified and identified as filamin C (FLN c), which is an actin-binding protein. TIM showed similar thermal and pH stability with better digestion resistance compared with FLN c. The result of the surface plasmon resonance (SPR) experiment demonstrated the infinity of anti-TIM polyclonal antibody (pAb) to both TIM and FLN c. Five linear and 3 conformational epitopes of TIM, as well as 9 linear and 10 conformational epitopes of FLN c, were mapped by phage display. Epitopes of TIM and FLN c demonstrated the sharing of certain residues; the occurrence of common epitopes in the two allergens accounts for their cross-reactivity.
磷酸丙糖异构酶（TIM）是糖酵解中的关键酶，已被确定为海产品中的一种过敏原。在本研究中，从淡水小龙虾（克氏原螯虾）肌肉中纯化出分子量为28 kDa的TIM。一种与TIM具有IgG/IgE交叉反应性的90 kDa蛋白质被纯化并鉴定为细丝蛋白C（FLN c），它是一种肌动蛋白结合蛋白。与FLN c相比，TIM表现出相似的热稳定性和pH稳定性，且具有更好的抗消化性。表面等离子体共振（SPR）实验结果表明，抗TIM多克隆抗体（pAb）对TIM和FLN c均具有无限亲和力。通过噬菌体展示技术定位了TIM的5个线性表位和3个构象表位，以及FLN c的9个线性表位和10个构象表位。TIM和FLN c的表位显示出某些残基的共享；两种过敏原中共同表位的出现解释了它们的交叉反应性。
