Purification and Characterization of Protamine, the Allergen from the Milt of Large Yellow Croaker (Pseudosciaena crocea), and Its Components

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• 作者：Yan-Yun Liu ; Xiao-Feng Chen ; Jia-Wei Hu ; Zhong-Wei Chen ; Ling-Jing Zhang ; Min-Jie Cao ; Guang-Ming Liu
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：2016
• 出版时间：March 9, 2016
• 年：2016
• 卷：64
• 期：9
• 页码：1999-2011
• 全文大小：660K
• ISSN：1520-5118

文摘

The protamine in fish milt can cause anaphylaxis in humans. To determine the allergen in the milt of large yellow croaker (Pseudosciaena crocea), crude extracts were incubated with sera from allergic patients. The results showed that a 12 kDa multicomponent protein was the major allergen in the milt of large yellow croaker. The multicomponent protein was purified, and physicochemical characterization showed that it was a glycoprotein, highly stable in acid–alkali conditions, and weakly retained immunoglobulin E (IgE)-binding activity at high temperatures. Separation and immunoreactivity analysis of the components of the multicomponent protein showed that it had six components, and component 5 had the strongest IgE-binding activity with patient sera. N-terminal sequencing confirmed the multicomponent protein was protamine. Following analysis of protamine from different fish by reversed-phase liquid chromatography and circular dichroism spectra, the protamines from different fish were found to have a similar secondary structure, although their components were different.