Page 94 - 《食品科学技术学报》2020年第5期

第 38 卷 第 5 期摇 摇 摇 摇 摇 摇 王郅媛等: 环核苷酸磷酸二酯酶 PDE9A 的体外表达、纯化与酶活特性分析 8 9

将相对酶活力提高 71郾 8% ,说明 PDE9A 具有一定 (24):10304 - 10313.
的金属离子依赖性且活性中心可能有半胱氨酸,另 [7]摇 TSAI E J, KASS D A. Cyclic GMP signaling in cardio鄄
外反应体系的缓冲液会影响 Mg 2 + 和 DTT 对蛋白质 vascular pathophysiology and therapeutics [ J]. Pharma鄄
cology and Therapeutics, 2009, 122(3):216 - 238.
活性的调节。
[8]摇 FISHER D A,SMITH J F,PILLAR J S, et al. Isolation
and characterization of PDE9A, a novel human cGMP鄄
3摇 结摇 论 specific phosphodiesterase[J]. J. Biol. Chem, 1998, 273
(3):570 - 577.
本研究通过 IPTG 诱导,使 PDE9A 蛋白在大肠 [9]摇 FERN魣NDEZ D, ROSENBROCK H, KROKER K S. In鄄
杆菌中大量异源表达,1 L 培养基可获得 3郾 125 g 蛋 hibition of PDE2A, but not PDE9A, modulates presynap鄄
白表达菌体。 采用多种纯化手段对目的蛋白进行纯 tic short鄄term plasticity measured by paired鄄pulse facilita鄄
tion in the CA1 region of the hippocampus[J]. Synapse,
化,1 g 表达菌体可纯化得到 8郾 9 mg 高纯度蛋白质。
2015, 69(10):484 - 496.
通过高效液相色谱技术检测,制备的 PDE9A 具有较
[10]摇 DHINGRA A,TUMMALA S R,LYUBARSKY A, et al.
2 + 和 DTT 能够显著提高
高的催化活性。 此外,Mg
PDE9A is expressed in the inner retina and contributes
2 + 和
PDE9A 活性,但反应体系的缓冲液会影响 Mg to the normal shape of the photopic ERG waveform[J].
DTT 对蛋白质活性的调节作用。 本研究最终制备得 Frontiers in Molecular Neuroscience, 2014, 7(60):1 -10.
到高纯度、高催化活性的 PDE9A 蛋白,可作为筛选 [11]摇 ALMEIDA C B,TRAINA F,LANARO C, et al. Locali鄄
zation of cyclic guanosine 3忆, 5忆鄄monophosphate鄄hydro鄄
具有调节血糖等功效的天然活性物质的靶点。
lyzing phosphodiesterase type 9 in rat brain by nonradio鄄
active in situ hydridization [ J]. Methods in Molecular
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[12]摇 KROKER K S, MATHIS C, MARTI A, et al. PDE9A
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