Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [57Fe2(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

doi:10.1021/jacs.5b03270

Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [⁵⁷Fe₂(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

Chemistry

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Abstract

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, S_8, 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 H_ox-CO state of [2⁵⁷Fe]_H and [4⁵⁷Fe-4S]_H HydA1 was characterized by Mössbauer, HYSCORE, ENDOR, and nuclear resonance vibrational spectroscopy. (Chemical Equation Presented).

Original language	English (US)
Pages (from-to)	8998-9005
Number of pages	8
Journal	Journal of the American Chemical Society
Volume	137
Issue number	28
DOIs	https://doi.org/10.1021/jacs.5b03270
State	Published - Jul 22 2015

ASJC Scopus subject areas

Catalysis
Chemistry(all)
Biochemistry
Colloid and Surface Chemistry

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10.1021/jacs.5b03270

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@article{d5627835a32541adbe023d1376d1c35d,

title = "Spectroscopic Investigations of [FeFe] Hydrogenase Maturated with [57Fe2(adt)(CN)2(CO)4]2-",

abstract = "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 M{\"o}ssbauer, HYSCORE, ENDOR, and nuclear resonance vibrational spectroscopy. (Chemical Equation Presented).",

author = "Ryan Gilbert-Wilson and Siebel, {Judith F.} and Agnieszka Adamska-Venkatesh and Pham, {Cindy C.} and Edward Reijerse and Hongxin Wang and Cramer, {Stephen P.} and Wolfgang Lubitz and Rauchfuss, {Thomas B.}",

note = "Publisher Copyright: {\textcopyright} 2015 American Chemical Society.",

year = "2015",

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doi = "10.1021/jacs.5b03270",

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AU - Gilbert-Wilson, Ryan

AU - Siebel, Judith F.

AU - Adamska-Venkatesh, Agnieszka

AU - Pham, Cindy C.

AU - Reijerse, Edward

AU - Wang, Hongxin

AU - Cramer, Stephen P.

AU - Lubitz, Wolfgang

AU - Rauchfuss, Thomas B.

PY - 2015/7/22

Y1 - 2015/7/22

N2 - 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. (Chemical Equation Presented).

AB - 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. (Chemical Equation Presented).

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