Characterization and in vitro interaction study of a [NiFe] hydrogenase large subunit from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1

Daisuke Sasaki, Satoshi Watanabe, Tamotsu Kanai, Haruyuki Atomi, Tadayuki Imanaka, Kunio Miki

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)192-196
Number of pages5
JournalBiochemical and biophysical research communications
Volume417
Issue number1
DOIs
Publication statusPublished - 2012 Jan 6
Externally publishedYes

Keywords

  • HypA
  • HypC
  • In vitro interaction
  • The large subunit HyhL
  • Thermococcus kodakarensis KOD1
  • [NiFe] hydrogenase maturation

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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