Heterologous expression and molecular characterization of the
NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus
denitrificans

J 2009

Heterologous expression and molecular characterization of the NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus denitrificans

TESAŘÍK, Radek, Vojtěch SEDLÁČEK, Jana PLOCKOVÁ, Michaela WIMMEROVÁ, Jaroslav TURÁNEK et. al.

Základní údaje

Originální název

Heterologous expression and molecular characterization of the NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus denitrificans

Název česky

Heterologni exprese a molekularni charakterizace NAD(P)H:acceptor oxidoreductasy (FerB) z Paracoccus denitrificans

Autoři

TESAŘÍK, Radek (203 Česká republika), Vojtěch SEDLÁČEK (203 Česká republika), Jana PLOCKOVÁ (203 Česká republika), Michaela WIMMEROVÁ (203 Česká republika), Jaroslav TURÁNEK (203 Česká republika) a Igor KUČERA (203 Česká republika, garant)

Vydání

Protein Expression and Purification, Elsevier, 2009, 1046-5928

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 1.563

Kód RIV

RIV/00216224:14310/09:00029452

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000272103000016

Klíčová slova anglicky

Quinone reductase; Protein expression in E. coli; His-tag fusion; Light scattering; Cross-linking; FT-IR spectrometry; Differential scanning calorimetry; Thermal inactivation

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 29. 3. 2010 11:04, prof. RNDr. Michaela Wimmerová, Ph.D.

Anotace

ORIG CZ

V originále

FerB is a flavoenzyme capable of reducing quinones, ferric complexes and chromate. Its expression in Escherichia coli as a hexahistidine fusion resulted in a functional product only when the tag was placed on the C-terminus. The molecular mass values estimated by gel permeation chromatography were compatible with the existence of either dimer or trimer, whereas the light scattering data, together with cross-linking experiments which yielded exclusively monomer and dimer bands on dodecyl sulfate-polyacrylamide gels, strongly supported a dimeric nature of both native and tagged form of FerB. These two proteins also exhibited almost identical secondary structures as judged by Fourier transform infra red spectrometry. The presence of tag, however, shifted the thermal denaturation curve towards lower temperatures and decreased the thermoresistance of enzyme activity. Despite somewhat lower thermal stability, the fusion protein is considered a better candidate for crystallization than the wild-type one due to a more negative value of its second optical virial coefficient.

Česky

Heterologni exprese a molekularni charakterizace NAD(P)H:acceptor oxidoreductasy (FerB) z Paracoccus denitrificans

Návaznosti

GA525/07/1069, projekt VaV

Název: Struktura, funkce a regulace FerB, širokospecifické bakteriální oxidoreduktasy s možným významem pro ekotechnologii

Investor: Grantová agentura ČR, Struktura, funkce a regulace FerB, širokospecifické bakteriální oxidoreduktasy s možným významem pro ekotechnologii

MSM0021622413, záměr

Název: Proteiny v metabolismu a při interakci organismů s prostředím

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Proteiny v metabolismu a při interakci organismů s prostředím

Citovat

TESAŘÍK, Radek, Vojtěch SEDLÁČEK, Jana PLOCKOVÁ, Michaela WIMMEROVÁ, Jaroslav TURÁNEK a Igor KUČERA. Heterologous expression and molecular characterization of the NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus denitrificans. Protein Expression and Purification. Elsevier, 2009, roč. 68, č. 2, s. 233-238. ISSN 1046-5928.

@article{842448,
   author = {Tesařík, Radek and Sedláček, Vojtěch and Plocková, Jana and Wimmerová, Michaela and Turánek, Jaroslav and Kučera, Igor},
   article_number = {2},
   keywords = {Quinone reductase; Protein expression in E. coli; His-tag fusion; Light scattering; Cross-linking; FT-IR spectrometry; Differential scanning calorimetry; Thermal inactivation},
   language = {eng},
   issn = {1046-5928},
   journal = {Protein Expression and Purification},
   title = {Heterologous expression and molecular characterization of the NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus denitrificans},
   volume = {68},
   year = {2009}
}

TY  - JOUR
ID  - 842448
AU  - Tesařík, Radek - Sedláček, Vojtěch - Plocková, Jana - Wimmerová, Michaela - Turánek, Jaroslav - Kučera, Igor
PY  - 2009
TI  - Heterologous expression and molecular characterization of the NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus denitrificans
JF  - Protein Expression and Purification
VL  - 68
IS  - 2
SP  - 233-238
EP  - 233-238
PB  - Elsevier
SN  - 10465928
KW  - Quinone reductase
KW  - Protein expression in E. coli
KW  - His-tag fusion
KW  - Light scattering
KW  - Cross-linking
KW  - FT-IR spectrometry
KW  - Differential scanning calorimetry
KW  - Thermal inactivation
N2  - FerB is a flavoenzyme capable of reducing quinones, ferric complexes and chromate. Its expression in Escherichia coli as a hexahistidine fusion resulted in a functional product only when the tag was placed on the C-terminus. The molecular mass values estimated by gel permeation chromatography were compatible with the existence of either dimer or trimer, whereas the light scattering data, together with cross-linking experiments which yielded exclusively monomer and dimer bands on dodecyl sulfate-polyacrylamide gels, strongly supported a dimeric nature of both native and tagged form of FerB. These two proteins also exhibited almost identical secondary structures as judged by Fourier transform infra red spectrometry. The presence of tag, however, shifted the thermal denaturation curve towards lower temperatures and decreased the thermoresistance of enzyme activity. Despite somewhat lower thermal stability, the fusion protein is considered a better candidate for crystallization than the wild-type one due to a more negative value of its second optical virial coefficient.
ER  -

TESAŘÍK, Radek, Vojtěch SEDLÁČEK, Jana PLOCKOVÁ, Michaela WIMMEROVÁ, Jaroslav TURÁNEK a Igor KUČERA. Heterologous expression and molecular characterization of the NAD(P)H:acceptor oxidoreductase (FerB) of Paracoccus denitrificans. \textit{Protein Expression and Purification}. Elsevier, 2009, roč.~68, č.~2, s.~233-238. ISSN~1046-5928.

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