Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [~(57)Fe_2(adt)(CN)_2(CO)_4]~(2-)

Ryan Gilbert-Wilson; Judith F. Siebel; Agnieszka Adamska-Venkatesh; Cindy C. Pham; Edward Reijerse; Hongxin Wang; Stephen P. Cramer; Wolfgang Lubitz; Thomas B. Rauchfuss

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Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [~(57)Fe_2(adt)(CN)_2(CO)_4]~(2-)

机译：[〜（57）Fe_2（adt）（CN）_2（CO）_4]〜（2-）饱和的[FeFe]氢化酶的光谱研究

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The preparation and spectroscopic characterization of a CO-inhibited [FeFe] hydrogenase with a selectively ~(57)Fe-labeled binuclear subsite is described. The precursor [~(57)Fe_2(adt)(CN)_2(CO)_4]~(2-) was synthesized from the ~(57)Fe metal, S_8, CO, (NEt_4)CN, NH_4Cl, and CH_2O. (Et_4N)_2[~(57)Fe_2(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 ~(57)Fe enrichment of the [4Fe-4S]_H cluster was realized by reconstirution with ~(57)FeCl_3 and Na_2S. The H_(OX)-CO state of [2~(57)Fe]_H and [4~(57)Fe-4S]_H HydA1 was characterized by Mossbauer, HYSCORE, ENDOR, and nuclear resonance vibrational spectroscopy.

机译：描述了具有选择性〜（57）Fe标记的双核亚位点的CO抑制的[FeFe]氢化酶的制备和光谱表征。由〜（57）Fe金属，S_8，CO，（NEt_4）CN，NH_4Cl和CH_2O合成前体[〜（57）Fe_2（adt）（CN）_2（CO）_4]〜（2-）。然后使用（Et_4N）_2 [〜（57）Fe_2（adt）（CN）_2（CO）_4]使莱茵衣藻的[FeFe]氢化酶HydA1成熟，以产生在[2Fe]处选择性标记的酶_H子集群。通过〜（57）FeCl_3和Na_2S的重构实现了[4Fe-4S] _H团簇的〜（57）Fe富集。 [2〜（57）Fe] _H和[4〜（57）Fe-4S] _H HydA1的H_（OX）-CO状态通过Mossbauer，HYSCORE，ENDOR和核共振振动光谱法表征。

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来源
《Journal of the American Chemical Society》 |2015年第28期|8998-9005|共8页
作者
Ryan Gilbert-Wilson; Judith F. Siebel; Agnieszka Adamska-Venkatesh; Cindy C. Pham; Edward Reijerse; Hongxin Wang; Stephen P. Cramer; Wolfgang Lubitz; Thomas B. Rauchfuss;
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作者单位

School of Chemical Sciences, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States;

Max-Planck-Institut fuer Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Muelheim an der Ruhr, Germany;

Max-Planck-Institut fuer Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Muelheim an der Ruhr, Germany;

Department of Chemistry, University of California, Davis, California 95616, United States;

Max-Planck-Institut fuer Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Muelheim an der Ruhr, Germany;

Department of Chemistry, University of California, Davis, California 95616, United States,Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States;

Department of Chemistry, University of California, Davis, California 95616, United States ,Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, United States;

Max-Planck-Institut fuer Chemische Energiekonversion, Stiftstrasse 34-36, 45470 Muelheim an der Ruhr, Germany;

School of Chemical Sciences, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, United States;

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入库时间 2022-08-18 03:09:47

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Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [~(57)Fe_2(adt)(CN)_2(CO)_4]~(2-)

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