Wang Jin-Song, Zheng Zhong-Liang, Lei Jing, Pan Ji-Cheng, Zou Guo-Lin
State Key Laboratory of Virology, College of Life Sciences, Center of Nanoscience and Nanotechnology, Wuhan University, Wuhan, Hubei 430072, PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2009 Jul;153(3):268-74. doi: 10.1016/j.cbpb.2009.03.010. Epub 2009 Mar 31.
Arginine kinase (AK) plays an important role in cellular energy metabolism in invertebrate. The encoding AK gene from Shrimp Metapenaeus ensis (M. ensis) was cloned in prokaryotic expression plasmid pET-28a, and it was then expressed in Escherichia coil in dissoluble form. The recombinant protein was purified by following three chromatography steps in turn: CM-Cellulose cation-exchange, Sephacryl S-100HR gel filtrate and DEAE-Sepharose anion-exchange. The purified AK's apparent K(m) was 2.33+/-0.1 and 1.59+/-0.2 mM for ATP and l-arginine, respectively, while its optimum pH and temperature was 8.5 and 30 degrees C in the process of forward reaction, respectively. Phylogenetic analysis of cDNA-derived amino acid sequences for the AKs indicated a close affinity of M. ensis and another shrimp (Litopenaeus vannamei).
精氨酸激酶（AK）在无脊椎动物的细胞能量代谢中起着重要作用。从凡纳滨对虾（M. ensis）中编码AK的基因被克隆到原核表达质粒pET-28a中，然后以不溶性形式在大肠杆菌中表达。重组蛋白依次通过以下三步色谱法进行纯化：CM-纤维素阳离子交换、Sephacryl S-100HR凝胶过滤和DEAE-琼脂糖阴离子交换。纯化后的AK对ATP和L-精氨酸的表观K(m)分别为2.33±0.1和1.59±0.2 mM，而其在正向反应过程中的最适pH和温度分别为8.5和30℃。对AK的cDNA推导氨基酸序列进行系统发育分析表明，凡纳滨对虾与另一种虾（南美白对虾）具有密切的亲缘关系。
