| Purification and identification of a clotting protein from the hemolymph of Chinese shrimp (Fenneropenaeus chinensis) | |
Wang Baojie1,2; Peng Hongni; Liu Mei1; Jiang Keyong1; Zhang Guofan1; Wang Lei1; Wang, L
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| 2013-09-01 | |
| 发表期刊 | JOURNAL OF OCEAN UNIVERSITY OF CHINA
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| ISSN | 1672-5182 |
| 卷号 | 12期号:3页码:477-483 |
| 文章类型 | Article |
| 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.; 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. |
| 关键词 | Fenneropenaeus Chinensis Clotting Protein Purification Proteomic Identification Maldi-tof/tof Ms |
| 学科领域 | Oceanography |
| DOI | 10.1007/s11802-013-2026-y |
| URL | 查看原文 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS研究方向 | Oceanography |
| WOS类目 | Oceanography |
| WOS记录号 | WOS:000322258300020 |
| WOS关键词 | FRESH-WATER CRAYFISH ; MASS-SPECTROMETRY ; PENAEUS-MONODON ; TIGER SHRIMP ; PACIFASTACUS-LENIUSCULUS ; CLOTTABLE PROTEINS ; COAGULATION ; VANNAMEI ; CLONING ; CRUSTACEANS |
| WOS标题词 | Science & Technology ; Physical Sciences |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qdio.ac.cn/handle/337002/16716 |
| 专题 | 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. |
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