Unique ligand-protein interactions in a new truncated hemoglobin from Mycobacterium tuberculosis.
A new truncated hemoglobin (HbO) from Mycobacterium tuberculosis has been expressed and purified. Sequence alignment of HbO with other hemoglobins suggests that the proximal F8 residue is histidine and the distal E7 and the B10 positions are occupied by alanine and tyrosine, respectively. The highly conserved residue at the CD1 position, surprisingly, is tyrosine, making HbO the first exception in the hemoglobin family that does not contain phenylalanine at this position. Resonance Raman data suggest that a strong hydrogen bonding network, involving the B10 Tyr and the CD1 Tyr, stabilizes the heme-bound O2 and CO as evidenced by the relatively low frequency of the Fe-O2 stretching mode (559 cm(-1)) and the high frequency of the Fe-CO stretching mode (527 cm(-1)). The presence of this hydrogen bonding network is supported by mutagenesis studies with the B10 tyrosine or the CD1 tyrosine mutated to phenylalanine. Taken together, these data demonstrate a rigid and polar distal pocket in HbO, which is significantly different from that of HbN, the other hemoglobin from M. tuberculosis. The distinct features in the heme active site structures and the temporal expression patterns of HbO and HbN suggest that these two hemoglobins may have very different physiological functions.
Mukai
Masahiro
M
Department of Physiology and Biophysics, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
Savard
Pierre-Yves
PY
Ouellet
Hugues
H
Guertin
Michel
M
Yeh
Syun-Ru
SR
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D006454:Hemoglobins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006860:Hydrogen Bonding; D008024:Ligands; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013059:Spectrum Analysis, Raman; D054793:Truncated Hemoglobins
D001426:Bacterial Proteins; D017931:DNA Primers; D006454:Hemoglobins; D008024:Ligands; D054793:Truncated Hemoglobins; C451738:hemoglobin O, Mycobacterium tuberculosis