Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [(57)Fe2(adt)(CN)2(CO)4](2-).
Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [(57)Fe2(adt)(CN)2(CO)4](2-).
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DOI：
10.1021/jacs.5b03270
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发表时间：
2015-07-22
影响因子：
15
通讯作者：
Rauchfuss TB
中科院分区：
文献类型：
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作者：
Gilbert-Wilson R;Siebel JF;Adamska-Venkatesh A;Pham CC;Reijerse E;Wang H;Cramer SP;Lubitz W;Rauchfuss TB
本文报道了一种具有选择性57 Fe标记双核亚位点的CO抑制[FeFe]氢化酶的制备和光谱表征。以~（57）Fe金属、S_8、CO、（NEt_4）CN、NH_4Cl和CH_2O为原料，合成了[~（57）Fe_2（adt）（CN）_2（CO）_4]_2-。（Et 4 N）2[57 Fe 2（adt）（CN）2（CO）4]用于来自莱茵衣藻的[FeFe]氢化酶HydA 1的成熟，以产生在[2Fe]H亚簇处选择性标记的酶。[4Fe-4S]H团簇通过与57 FeCl 3和Na 2S的重组实现了57 Fe的互补富集。[257 Fe]H和[457 Fe-4S]H HydA 1的Hox-CO态用穆斯堡尔谱、HYSCORE谱、ENDOR谱和核共振振动谱进行了表征。
The preparation and spectroscopic characterization of a CO-inhibited [FeFe] hydrogenase with a selectively 57Fe-labeled binuclear subsite is described. The precursor [57Fe2(adt)(CN)2(CO)4]2– was synthesized from the 57Fe metal, S8, CO, (NEt4)CN, NH4Cl, and CH2O. (Et4N)2[57Fe2(adt)(CN)2(CO)4] was then used for the maturation of the [FeFe] hydrogenase HydA1 from Chlamydomonas reinhardtii, to yield the enzyme selectively labeled at the [2Fe]H subcluster. Complementary 57Fe enrichment of the [4Fe-4S]H cluster was realized by reconstitution with 57FeCl3 and Na2S. The Hox-CO state of [257Fe]H and [457Fe-4S]H HydA1 was characterized by Mössbauer, HYSCORE, ENDOR, and nuclear resonance vibrational spectroscopy.