Purification and Identification of a Clotting Protein from the Hemolymph of Chinese Shrimp （Fenneropenaeus chinensis）

• Published in: 中国海洋大学学报：英文版 no. 3; pp. 477 - 483
• Main Author: WANG Baojie PENG Hongni LIU Mei JIANG Keyong ZHANG Guofan WANG Lei
• Format: Journal Article
• Language: English
• Published: 2013
• Subjects: SDS-PAGE电泳; 中国对虾; 凝血机制; 基质辅助激光解吸; 层析纯化; 胰蛋白酶; 血淋巴; 阴离子交换色谱
• ISSN: 1672-5182; 1993-5021

The clotting protein （CP） plays important and diverse roles in crustaceans, such as coagulation and lipid transportation. A clotting protein was purified from the hemolymph of Chinese shrimp Fenneropenaeus chinensis （named as Fc-CP） with Q sepha- rose HP anion-exchange chromatography and phenyl sepharose HP hydrophobic interaction chromatography. Fc-CP was able to form stable clots in vitro in the presence of hemocyte lysate and Ca2~, suggesting that the clotting reaction is catalyzed by a Ca2＋-dependent transglutaminase in shrimp hemocytes. The molecular mass of Fc-CP was 380 kDa under non-reducing conditions and 190 kDa under reducing conditions as was determined with SDS-PAGE. CP exists as disulfide-linked homodimers and oligomers The N-terminal amino acid sequence of Fc-CP was identical to that of shrimps including Penaeus monodon, Farfantepenaeus paulensis and Litopenaeus vannamei; and similar to that of other decapods. The purified Fc-CP was digested with trypsin and veri- fied on an ABI 4700 matrix-assisted laser desorption/ionization tandem time-of-flight （MALDI-TOF/TOF） mass spectrometry. Our results will aid to better understanding the coagulation mechanism of shrimp hemolymph.