Novel FixL homologues in Chlamydomonas reinhardtii bind heme and O 2

U. M Narayana Murthy, Matt S A Wecker, Matthew C. Posewitz, Marie Alda Gilles-Gonzalez, Maria L. Ghirardi

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Genome inspection revealed nine putative heme-binding, FixL-homologous proteins in Chlamydomonas reinhardtii. The heme-binding domains from two of these proteins, FXL1 and FXL5 were cloned, expressed in Escherichia coli, purified and characterized. The recombinant FXL1 and FXL5 domains stained positively for heme, while mutations in the putative ligand-binding histidine FXL1-H200S and FXL5-H200S resulted in loss of heme binding. The FXL1 and FXL5 [Fe(II), bound O2] had Soret absorption maxima around 415 nm, and weaker absorptions at longer wavelengths, in concurrence with the literature. Ligand-binding measurements showed that FXL1 and FXL5 bind O2 with moderate affinity, 135 and 222 μM, respectively. This suggests that Chlamydomonas may use the FXL proteins in O2-sensing mechanisms analogous to that reported in nitrogen-fixing bacteria to regulate gene expression.

Original languageEnglish (US)
Pages (from-to)4282-4288
Number of pages7
JournalFEBS Letters
Volume586
Issue number24
DOIs
StatePublished - Dec 14 2012

Fingerprint

Chlamydomonas reinhardtii
Heme
Ligands
Chlamydomonas
Proteins
Histidine
Gene expression
Escherichia coli
Bacteria
Nitrogen
Genes
Inspection
Genome
Gene Expression
Wavelength
Mutation
heme O

Keywords

  • Chlamydomonas reinhardtii
  • FixL
  • Heme-binding
  • Oxygen-sensor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Genetics
  • Molecular Biology
  • Structural Biology

Cite this

Murthy, U. M. N., Wecker, M. S. A., Posewitz, M. C., Gilles-Gonzalez, M. A., & Ghirardi, M. L. (2012). Novel FixL homologues in Chlamydomonas reinhardtii bind heme and O 2 FEBS Letters, 586(24), 4282-4288. https://doi.org/10.1016/j.febslet.2012.06.052

Novel FixL homologues in Chlamydomonas reinhardtii bind heme and O 2 . / Murthy, U. M Narayana; Wecker, Matt S A; Posewitz, Matthew C.; Gilles-Gonzalez, Marie Alda; Ghirardi, Maria L.

In: FEBS Letters, Vol. 586, No. 24, 14.12.2012, p. 4282-4288.

Research output: Contribution to journalArticle

Murthy, UMN, Wecker, MSA, Posewitz, MC, Gilles-Gonzalez, MA & Ghirardi, ML 2012, 'Novel FixL homologues in Chlamydomonas reinhardtii bind heme and O 2 ', FEBS Letters, vol. 586, no. 24, pp. 4282-4288. https://doi.org/10.1016/j.febslet.2012.06.052
Murthy, U. M Narayana ; Wecker, Matt S A ; Posewitz, Matthew C. ; Gilles-Gonzalez, Marie Alda ; Ghirardi, Maria L. / Novel FixL homologues in Chlamydomonas reinhardtii bind heme and O 2 In: FEBS Letters. 2012 ; Vol. 586, No. 24. pp. 4282-4288.
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