Characterization and in vitro Interaction Study of a [NiFe] Hydrogenase Large Subunit from the Hyperthermophilic Archaeon Thermococcus kodakarensis KOD1
Characterization and in vitro Interaction Study of a [NiFe] Hydrogenase Large Subunit from the Hyperthermophilic Archaeon Thermococcus kodakarensis KOD1
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超嗜热古细菌 Thermococcus kodakarensis KOD1 的 [NiFe] 氢化酶大亚基的表征和体外相互作用研究
DOI：
10.1016/j.bbrc.2011.11.083
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
and K. Miki
中科院分区：
文献类型：
--
作者：
D. Sasaki;S. Watanabe;T. Kanai;H. Atomi;T. Imanaka;and K. Miki
[NiFe]氢化酶的大亚基含有NiFe（CN）2（CO）簇。成熟蛋白HypA、B、C、D、E和F是NiFe簇生物合成所需的。虽然成熟机制迄今已被深入研究，很少有人知道Hyp蛋白和大亚基的[NiFe]氢化酶之间的相互作用。在这项研究中，我们已经纯化和表征胞质[NiFe]氢化酶大亚基从Thermococcus kodakarensis（Tk-HyhL）。Tk-HyhL在单体和二聚体形式之间平衡存在。体外相互作用分析表明，Tk-HyhL单体与Tk-HypA形成紧密复合物，与Tk-HypC弱相互作用。未检测到预期的三元复合物形成。这些意见反映了多样性的机制，镍插入[NiFe]氢化酶成熟取决于生物体。
The large subunit of the [NiFe] hydrogenases harbors a NiFe(CN)2(CO) cluster. Maturation proteins HypA, B, C, D, E, and F are required for the NiFe cluster biosynthesis. While the maturation machinery has been hitherto studied intensively, little is known about interactions between the Hyp proteins and the large subunit of the [NiFe] hydrogenase. In this study, we have purified and characterized the cytosolic [NiFe] hydrogenase large subunit HyhL from Thermococcus kodakarensis (Tk-HyhL). Tk-HyhL exists in equilibrium between monomeric and dimeric forms. In vitro interaction analyses showed that Tk-HyhL monomer forms a tight complex with Tk-HypA and weakly interacts with Tk-HypC. The expected ternary complex formation was not detected. These observations reflect a diversity in the mechanism of Ni insertion in [NiFe] hydrogenase maturation depending on the organism.