TY - JOUR
T1 - Characterization and in vitro interaction study of a [NiFe] hydrogenase large subunit from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1
AU - Sasaki, Daisuke
AU - Watanabe, Satoshi
AU - Kanai, Tamotsu
AU - Atomi, Haruyuki
AU - Imanaka, Tadayuki
AU - Miki, Kunio
N1 - Funding Information:
This work was partly supported by Grants-in-Aid for Scientific Research (A) (to KM) and by the Global COE program “ International Center for Integrated Research and Advanced Education in Materials Science ” (to DS) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan. The metal analysis of Tk-HyhL was performed by TORAY Research Center Inc.
PY - 2012/1/6
Y1 - 2012/1/6
N2 - The large subunit of the [NiFe] hydrogenases harbors a NiFe(CN) 2(CO) cluster. Maturation proteins HypA, B, C, D, E, and F are required for the NiFe cluster biosynthesis. While the maturation machinery has been hitherto studied intensively, little is known about interactions between the Hyp proteins and the large subunit of the [NiFe] hydrogenase. In this study, we have purified and characterized the cytosolic [NiFe] hydrogenase large subunit HyhL from Thermococcus kodakarensis (Tk-HyhL). Tk-HyhL exists in equilibrium between monomeric and dimeric forms. In vitro interaction analyses showed that Tk-HyhL monomer forms a tight complex with Tk-HypA and weakly interacts with Tk-HypC. The expected ternary complex formation was not detected. These observations reflect a diversity in the mechanism of Ni insertion in [NiFe] hydrogenase maturation depending on the organism.
AB - The large subunit of the [NiFe] hydrogenases harbors a NiFe(CN) 2(CO) cluster. Maturation proteins HypA, B, C, D, E, and F are required for the NiFe cluster biosynthesis. While the maturation machinery has been hitherto studied intensively, little is known about interactions between the Hyp proteins and the large subunit of the [NiFe] hydrogenase. In this study, we have purified and characterized the cytosolic [NiFe] hydrogenase large subunit HyhL from Thermococcus kodakarensis (Tk-HyhL). Tk-HyhL exists in equilibrium between monomeric and dimeric forms. In vitro interaction analyses showed that Tk-HyhL monomer forms a tight complex with Tk-HypA and weakly interacts with Tk-HypC. The expected ternary complex formation was not detected. These observations reflect a diversity in the mechanism of Ni insertion in [NiFe] hydrogenase maturation depending on the organism.
KW - HypA
KW - HypC
KW - In vitro interaction
KW - The large subunit HyhL
KW - Thermococcus kodakarensis KOD1
KW - [NiFe] hydrogenase maturation
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U2 - 10.1016/j.bbrc.2011.11.083
DO - 10.1016/j.bbrc.2011.11.083
M3 - Article
C2 - 22138400
AN - SCOPUS:84855793147
SN - 0006-291X
VL - 417
SP - 192
EP - 196
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -