A four-domain Kunitz-type proteinase inhibitor from Solen grandis is implicated in immune response

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• 作者：Xiumei Wei^a ; Jialong Yang^b ; ^{jlyang@yic.ac.cn} ; Jianmin Yang^a ; Xiangquan Liu^a ; ^{lxq6808@163.com} ; Meijun Liu^a ; Dinglong Yang^a ; Jie Xu^a ; Xiaoke Hu^b
• 关键词：Solen grandis ; Kunitz-type proteinase inhibitor ; Innate immunity ; Real-time PCR ; Inhibitory activity
• 刊名：Fish & Shellfish Immunology
• 出版年：2012
• 出版时间：December, 2012
• 年：2012
• 卷：33
• 期：6
• 页码：1276-1284
• 全文大小：1387 K

文摘

Serine proteinase inhibitor (SPI) serves as a negative regulator in immune signal pathway by restraining the activities of serine proteinase (SP) and plays an essential role in the innate immunity. In the present study, a Kunitz-type SPI was identified from the mollusk razor clam Solen grandis (designated as SgKunitz). The full-length cDNA of SgKunitz was of 1284?bp, containing an open reading frame (ORF) of 768?bp. The ORF encoded four Kunitz domains, and their amino acids were well conserved when compared with those in other Kunitz-type SPIs, especially the six cysteines involved in forming of three disulfide bridges in each domain. In addition, the tertiary structure of all the four domains adopted a typical model of Kunitz-type SPI family, indicating SgKunitz was a new member of Kunitz-type SPI superfamily. The mRNA transcripts of SgKunitz were detected in all tested tissues of razor clam, including muscle, mantle, gonad, gill, hepatopancreas and hemocytes, and with the highest expression level in gill. When the razor clams were stimulated by LPS, PGN or ¦Â-1, 3-glucan, the expression level of SgKunitz mRNA in hemocytes was significantly up-regulated (P?<?0.01), suggesting SgKunitz might involved in the processes of inhibiting the activity of SPs during the immune responses triggered by various pathogens. Furthermore, the recombinant protein of SgKunitz could effectively inhibit the activities of SP trypsin and chymotrypsin in?vitro. The present results suggested SgKunitz could serve as an inhibitor of SP involving in the immune response of S. grandis, and provided helpful evidences to understand the regulation mechanism of immune signal pathway in mollusk.