Selenite reduction by the thioredoxin system: Kinetics and identification of protein-bound selenide

Takashi Tamura; Kumi Sato; Kentaro Komori; Takeshi Imai; Mitsuhiko Kuwahara; Takahiro Okugochi; Hisaaki Mihara; Nobuyoshi Esaki; Kenji Inagaki

doi:10.1271/bbb.100847

Selenite reduction by the thioredoxin system: Kinetics and identification of protein-bound selenide

Takashi Tamura, Kumi Sato, Kentaro Komori, Takeshi Imai, Mitsuhiko Kuwahara, Takahiro Okugochi, Hisaaki Mihara, Nobuyoshi Esaki, Kenji Inagaki

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

Selenite (SeO₃^2-) assimilation into a bacterial selenoprotein depends on thioredoxin (trx) reductase in Esherichia coli, but the molecular mechanism has not been elucidated. The mineral-oil overlay method made it possible to carry out anaerobic enzyme assay, which demonstrated an initial lag-phase followed by timedependent steady NADPH consumption with a positive cooperativity toward selenite and trx. SDS-PAGE/autoradiography using 75Se-labeled selenite as substrate revealed the formation of trx-bound selenium in the reaction mixture. The protein-bound selenium has metabolic significance in being stabilized in the divalent state, and it also produced the selenopersulfide (-S-SeH) form by the catalysis of E. coli trx reductase (TrxB).

Original language	English
Pages (from-to)	1184-1187
Number of pages	4
Journal	Bioscience, Biotechnology and Biochemistry
Volume	75
Issue number	6
DOIs	https://doi.org/10.1271/bbb.100847
Publication status	Published - 2011

Keywords

Anaerobic enzyme assay
Escherichia coli
Selenite
Selenopersulfide
Thioredoxin reductase

ASJC Scopus subject areas

Biotechnology
Analytical Chemistry
Biochemistry
Applied Microbiology and Biotechnology
Molecular Biology
Organic Chemistry

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10.1271/bbb.100847

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title = "Selenite reduction by the thioredoxin system: Kinetics and identification of protein-bound selenide",

abstract = "Selenite (SeO32-) assimilation into a bacterial selenoprotein depends on thioredoxin (trx) reductase in Esherichia coli, but the molecular mechanism has not been elucidated. The mineral-oil overlay method made it possible to carry out anaerobic enzyme assay, which demonstrated an initial lag-phase followed by timedependent steady NADPH consumption with a positive cooperativity toward selenite and trx. SDS-PAGE/autoradiography using 75Se-labeled selenite as substrate revealed the formation of trx-bound selenium in the reaction mixture. The protein-bound selenium has metabolic significance in being stabilized in the divalent state, and it also produced the selenopersulfide (-S-SeH) form by the catalysis of E. coli trx reductase (TrxB).",

keywords = "Anaerobic enzyme assay, Escherichia coli, Selenite, Selenopersulfide, Thioredoxin reductase",

author = "Takashi Tamura and Kumi Sato and Kentaro Komori and Takeshi Imai and Mitsuhiko Kuwahara and Takahiro Okugochi and Hisaaki Mihara and Nobuyoshi Esaki and Kenji Inagaki",

year = "2011",

doi = "10.1271/bbb.100847",

language = "English",

volume = "75",

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T2 - Kinetics and identification of protein-bound selenide

AU - Tamura, Takashi

AU - Sato, Kumi

AU - Komori, Kentaro

AU - Imai, Takeshi

AU - Kuwahara, Mitsuhiko

AU - Okugochi, Takahiro

AU - Mihara, Hisaaki

AU - Esaki, Nobuyoshi

AU - Inagaki, Kenji

PY - 2011

Y1 - 2011

N2 - Selenite (SeO32-) assimilation into a bacterial selenoprotein depends on thioredoxin (trx) reductase in Esherichia coli, but the molecular mechanism has not been elucidated. The mineral-oil overlay method made it possible to carry out anaerobic enzyme assay, which demonstrated an initial lag-phase followed by timedependent steady NADPH consumption with a positive cooperativity toward selenite and trx. SDS-PAGE/autoradiography using 75Se-labeled selenite as substrate revealed the formation of trx-bound selenium in the reaction mixture. The protein-bound selenium has metabolic significance in being stabilized in the divalent state, and it also produced the selenopersulfide (-S-SeH) form by the catalysis of E. coli trx reductase (TrxB).

AB - Selenite (SeO32-) assimilation into a bacterial selenoprotein depends on thioredoxin (trx) reductase in Esherichia coli, but the molecular mechanism has not been elucidated. The mineral-oil overlay method made it possible to carry out anaerobic enzyme assay, which demonstrated an initial lag-phase followed by timedependent steady NADPH consumption with a positive cooperativity toward selenite and trx. SDS-PAGE/autoradiography using 75Se-labeled selenite as substrate revealed the formation of trx-bound selenium in the reaction mixture. The protein-bound selenium has metabolic significance in being stabilized in the divalent state, and it also produced the selenopersulfide (-S-SeH) form by the catalysis of E. coli trx reductase (TrxB).

KW - Anaerobic enzyme assay

KW - Escherichia coli

KW - Selenite

KW - Selenopersulfide

KW - Thioredoxin reductase

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JO - Bioscience, Biotechnology and Biochemistry

JF - Bioscience, Biotechnology and Biochemistry

IS - 6

ER -

Selenite reduction by the thioredoxin system: Kinetics and identification of protein-bound selenide

Abstract

Keywords

ASJC Scopus subject areas

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