Zhou G, Parthasarathy G, Somasundaram T, Ables A, Roy L, Strong S J, Ellington W R, Chapman M S
Department of Chemistry, Florida State University, Tallahassee 32306-3015, USA.
Protein Sci. 1997 Feb;6(2):444-9. doi: 10.1002/pro.5560060222.
Phosphagen kinases catalyze the reversible transfer of a phosphoryl group between guanidino phosphate compounds and ADP, thereby regenerating ATP during bursts of cellular activity. Large quantities of highly pure arginine kinase (EC 2.7.3.3), the phosphagen kinase present in arthropods, have been isolated from E. coli, into which the cDNA for the horseshoe crab enzyme had been cloned. Purification involves size exclusion and anion exchange chromatographies applied in the denatured and refolded states. The recombinant enzyme has been crystallized as a transition state analog complex. Near complete native diffraction data have been collected to 1.86 A resolution. Substitution of a recombinant source for a natural one, improvement in the purification, and data collection at cryo temperatures have all yielded significant improvements in diffraction.
磷酸原激酶催化胍基磷酸化合物与ADP之间磷酸基团的可逆转移，从而在细胞活动爆发期间再生ATP。大量高纯度的精氨酸激酶（EC 2.7.3.3），即节肢动物中存在的磷酸原激酶，已从大肠杆菌中分离出来，其中已克隆了鲎酶的cDNA。纯化涉及在变性和复性状态下应用的尺寸排阻色谱和阴离子交换色谱。重组酶已结晶为过渡态类似物复合物。已收集到分辨率为1.86 Å的近乎完整的天然衍射数据。用重组来源替代天然来源、改进纯化方法以及在低温下进行数据收集，都在衍射方面取得了显著改进。
