TY - JOUR
T1 - Novel FixL homologues in Chlamydomonas reinhardtii bind heme and O 2
AU - Murthy, U. M Narayana
AU - Wecker, Matt S A
AU - Posewitz, Matthew C.
AU - Gilles-Gonzalez, Marie Alda
AU - Ghirardi, Maria L.
N1 - Funding Information:
This work was supported by the a grant from the Chemical Sciences, Geosciences and Biological Sciences Program, Division of Energy Biosciences, Office of Science, U.S. Department of Energy.
PY - 2012/12/14
Y1 - 2012/12/14
N2 - 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.
AB - 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.
KW - Chlamydomonas reinhardtii
KW - FixL
KW - Heme-binding
KW - Oxygen-sensor
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U2 - 10.1016/j.febslet.2012.06.052
DO - 10.1016/j.febslet.2012.06.052
M3 - Article
C2 - 22801216
AN - SCOPUS:84870519743
SN - 0014-5793
VL - 586
SP - 4282
EP - 4288
JO - FEBS Letters
JF - FEBS Letters
IS - 24
ER -