Identification and characterization of PshBII, a second FA/FB-containing polypeptide in the photosynthetic reaction center of Heliobacterium modesticaldum

Steven P. Romberger, Christian Castro, Yili Sun, John H. Golbeck

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

All known Type I photosynthetic reaction centers harbor three [4Fe-4S] clusters named FX, FA and FB that function as terminal electron acceptors. We reported earlier that FA and FB in the homodimeric Type I reaction center from Heliobacterium modesticaldum reside on a loosely bound 54 amino acid protein named PshB. Time-resolved optical spectroscopy and low temperature EPR spectroscopy showed that on illumination, electrons were transferred from FX - to FA and FB at both cryogenic and room temperatures. Interestingly, the gene that codes for PshB, HM1_1462, is part of a predicted dicistronic operon that contains a second gene, named HM1_1461, which codes for a second ferredoxin-like protein with high sequence homology to PshB, including the two traditional [4Fe-4S] cluster binding motifs. RT-PCR results confirm that both genes are transcribed as a single transcript. We have cloned the HM1_1461 gene through PCR amplification of the H. modesticaldum chromosomal DNA and overexpressed the apoprotein in Escherichia coli. Reconstitution studies with inorganic reagents have shown that the holoprotein harbors ~8 iron and ~8 sulfide atoms in the form of two [4Fe-4S] clusters. Incubation of the reconstituted holoprotein with heliobacterial reaction center cores results in a charge-separated state characteristic of electron transfer past the FX cluster to the terminal [4Fe-4S] clusters FA and FB. These results suggest that the HM1_1461 product, which we have named PshBII, is capable of functioning in lieu of PshB (renamed PshBI) as an alternative terminal electron transfer protein. Thus, unlike PS I, to which PsaC is tightly bound, two loosely bound ferredoxins, PshBI and PshBII, are capable of interacting with the heliobacterial reaction center. The presence of two, loosely bound FA/FB proteins represents a significant shift in our understanding of structure-function relationships in Type I reaction centers.

Original languageEnglish (US)
Pages (from-to)293-303
Number of pages11
JournalPhotosynthesis research
Volume104
Issue number2
DOIs
StatePublished - Jun 1 2010

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Cell Biology
  • Biochemistry

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