Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)

	Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)
	Wang Baojie 1,2; Peng Hongni ; Liu Mei 1; Jiang Keyong 1; Zhang Guofan 1; Wang Lei 1; Wang, L
刊名	JOURNAL OF OCEAN UNIVERSITY OF CHINA
	2013-09-01
卷号	12 期号:3 页码:477-483
关键词	Fenneropenaeus chinensis clotting protein purification proteomic identification MALDI-TOF/TOF MS
ISSN号	1672-5182
通讯作者	Wang, L
中文摘要	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 sepharose 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 verified 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.
英文摘要	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 sepharose 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 verified 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.
学科主题	Oceanography
WOS标题词	Science & Technology ; Physical Sciences
类目[WOS]	Oceanography
研究领域[WOS]	Oceanography
关键词[WOS]	FRESH-WATER CRAYFISH ; MASS-SPECTROMETRY ; PENAEUS-MONODON ; TIGER SHRIMP ; PACIFASTACUS-LENIUSCULUS ; CLOTTABLE PROTEINS ; COAGULATION ; VANNAMEI ; CLONING ; CRUSTACEANS
收录类别	SCI
原文出处	10.1007/s11802-013-2026-y
语种	英语
WOS记录号	WOS:000322258300020
公开日期	2014-07-17
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/16716]
专题	海洋研究所_海洋生物技术研发中心
通讯作者	Wang, L
作者单位	1.Chinese Acad Sci, Inst Oceanl, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Wang Baojie,Peng Hongni,Liu Mei,et al. Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)[J]. JOURNAL OF OCEAN UNIVERSITY OF CHINA,2013,12(3):477-483.
APA	Wang Baojie.,Peng Hongni.,Liu Mei.,Jiang Keyong.,Zhang Guofan.,...&Wang, L.(2013).Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis).JOURNAL OF OCEAN UNIVERSITY OF CHINA,12(3),477-483.
MLA	Wang Baojie,et al."Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis)".JOURNAL OF OCEAN UNIVERSITY OF CHINA 12.3(2013):477-483.