Stabilization of iron-sulfur cluster FX by intra-subunit interactions unraveled by suppressor and second site-directed mutations in PsaB of Photosystem I

Ming Tao Zeng, Xiao Min Gong, Michael C.W. Evans, Nathan Nelson, Chanoch Carmeli*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Intra-subunit interactions in the environment of the iron-sulfur cluster FX in Photosystem I (PS I) of Synechocystis sp. PCC 6803 were studied by site-directed and second site suppressor mutations. In subunit PsaB, the cysteine ligand (C565) of FX and a conserved aspartate (D566) adjacent to C565 were modified. The resulting mutants D566E, C556S/D566E, C556H/D566E and C565H/D566E did not assemble PS I in the thylakoids of the cyanobacterium. Yet, this is the first report of cells of the second site-suppressor mutant (D566E/L416P) and of second site-directed mutant (C565S/D566E) in PsaB that could grow autotrophically in light and were found to assemble a stable functional PS I containing all three iron-sulfur centers, FX and FA/B. The newly resolved structure of PS I (PDB 1JB0) was used to interpret the functional interactions among the amino acid residues. It is suggested that the stability of FX is supported by a salt bridge formed between D566, which is adjacent to the cysteine ligand C565 of the iron-sulfur cluster located on loop hi, and R703 located at the start of loop jk. Hydrogen bond between R703 and D571 at the start of loop hi further stabilizes the arginine. Lengthening of the side by 1.2 Å chain in mutation D566E caused destabilization of FX. The extended side-chain was compensated for by the Fe-O, which is 0.3 Å shorter than the Fe-S bond resulting in stabilization of the FX in the double mutations C565S/D566E. The suppressor mutation D566E/L416P allowed greater freedom for the salt bridge E566-R703, thus relieving the pressure introduced by the D566E replacement and enabling the formation of FX. FX and R703 are therefore stabilized through short- and long-range interactions of the inter-helical loops between h-i, j-k and f-g, respectively.

Original languageEnglish
Pages (from-to)254-264
Number of pages11
JournalBiochimica et Biophysica Acta - Bioenergetics
Volume1556
Issue number2-3
DOIs
StatePublished - 2 Dec 2002

Keywords

  • Electron paramagnetic resonance
  • Electron transport
  • Iron-sulfur cluster
  • Photosystem I
  • Spectroscopy
  • Suppressor mutation

Fingerprint

Dive into the research topics of 'Stabilization of iron-sulfur cluster FX by intra-subunit interactions unraveled by suppressor and second site-directed mutations in PsaB of Photosystem I'. Together they form a unique fingerprint.

Cite this