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Purification and Primary Identification of Haemocyanin in the Chinese Shrimp, Fenneropenaeus Chinensis (Decapoda, Penaeoidea)

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[We here report the isolation of haemocyanin, the crustacean oxygen transporter, from the haemolymph of the Chinese shrimp, Fenneropenaeus chinensis. We employed a Superdex 75 column, and the purified protein was demonstrated as homogeneous by Native-PAGE. The molecular mass of the purified haemocyanin was estimated to be 450 kDa using SDS-PAGE. The subsequent 2-DE suggested that it would be composed of three different subunits of 75-76 kDa each. Primary structure determination was performed by LC-ESI-MS and the data obtained were compared with the NCBI database. The result showed that these three subunits are identical to a partial fragment of haemocyanin from Litopenaeus vannamei. This research could be relevant to obtain pure haemocyanin, and will be helpful for further study on the structure and function of this large molecule. L'isolation de l'hémocyanine, le transporteur d'oxygène des crustacés, à partir de l'hémolymphe de la crevette chinoise Fenneropenaeus chinensis a été effectué. Nous avons utilisé une colonne Superdex 75 et la protéine purifiée a été démontrée homogène par Native-PAGE. Le poids moléculaire de l'hémocyanine purifiée a été estimé être de 450 kDa par SDS-PAGE. Le subséquent 2-DE a suggéré qu'elle pourrait être composée de trois différentes sous-unités de 75-76 kDa chacune. La détermination de la structure primaire a été réalisée par LC-ESI-MS et les données obtenues ont été comparées avec la base de données NCBI. Les résultats ont montré que ces trois sous-unités sont identiques à un fragment partiel d'hémocyanine de Litopenaeus vannamei. Ces recherches pourraient permettre d'obtenir de l'hémocyanine pure et seront utiles pour de futures études sur la structure et la fonction de cette grosse molécule., We here report the isolation of haemocyanin, the crustacean oxygen transporter, from the haemolymph of the Chinese shrimp, Fenneropenaeus chinensis. We employed a Superdex 75 column, and the purified protein was demonstrated as homogeneous by Native-PAGE. The molecular mass of the purified haemocyanin was estimated to be 450 kDa using SDS-PAGE. The subsequent 2-DE suggested that it would be composed of three different subunits of 75-76 kDa each. Primary structure determination was performed by LC-ESI-MS and the data obtained were compared with the NCBI database. The result showed that these three subunits are identical to a partial fragment of haemocyanin from Litopenaeus vannamei. This research could be relevant to obtain pure haemocyanin, and will be helpful for further study on the structure and function of this large molecule. L'isolation de l'hémocyanine, le transporteur d'oxygène des crustacés, à partir de l'hémolymphe de la crevette chinoise Fenneropenaeus chinensis a été effectué. Nous avons utilisé une colonne Superdex 75 et la protéine purifiée a été démontrée homogène par Native-PAGE. Le poids moléculaire de l'hémocyanine purifiée a été estimé être de 450 kDa par SDS-PAGE. Le subséquent 2-DE a suggéré qu'elle pourrait être composée de trois différentes sous-unités de 75-76 kDa chacune. La détermination de la structure primaire a été réalisée par LC-ESI-MS et les données obtenues ont été comparées avec la base de données NCBI. Les résultats ont montré que ces trois sous-unités sont identiques à un fragment partiel d'hémocyanine de Litopenaeus vannamei. Ces recherches pourraient permettre d'obtenir de l'hémocyanine pure et seront utiles pour de futures études sur la structure et la fonction de cette grosse molécule.]

Affiliations: 1: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P. R. China, Graduate School, Chinese Academy of Sciences, Beijing 100039, P. R. China; 2: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P. R. China; 3: Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P. R. China;, Email: jhxiang@ms.qdio.ac.cn

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/content/journals/10.1163/156854008784771630
2008-07-01
2016-12-08

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