title: A cooperative oxygen-binding hemoglobin from Mycobacterium tuberculosis.
abstract: Two putative hemoglobin genes, glbN and glbO, were recently discovered in the complete genome sequence of Mycobacterium tuberculosis H37Rv. Here, we show that the glbN gene encodes a dimeric hemoglobin (HbN) that binds oxygen cooperatively with very high affinity (P(50) = 0.013 mmHg at 20 degrees C) because of a fast combination (25 microM(-1).s(-1)) and a slow dissociation (0.2 s(-1)) rate. Resonance Raman spectroscopy and ligand association/dissociation kinetic measurements, along with mutagenesis studies, reveal that the stabilization of the bound oxygen is achieved through a tyrosine at the B10 position in the distal pocket of the heme with a conformation that is unique among the globins. Physiological studies performed with Mycobacterium bovis bacillus Calmette-Guérin demonstrate that the expression of HbN is greatly enhanced during the stationary phase in aerobic cultures but not under conditions of limited oxygen availability. The results suggest that, physiologically, the primary role of HbN may be to protect the bacilli against reactive nitrogen species produced by the host macrophage.
authors.affiliation: Department of Biochemistry, Faculty of Sciences and Engineering, Laval University, Quebec, QC Canada G1K 7P4.
authors.forename: M
authors.initials: M
authors.lastname: Couture
authors.forename: S R
authors.initials: SR
authors.lastname: Yeh
authors.forename: B A
authors.initials: BA
authors.lastname: Wittenberg
authors.forename: J B
authors.initials: JB
authors.lastname: Wittenberg
authors.forename: Y
authors.initials: Y
authors.lastname: Ouellet
authors.forename: D L
authors.initials: DL
authors.lastname: Rousseau
authors.forename: M
authors.initials: M
authors.lastname: Guertin
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D006454:Hemoglobins / Q000737:chemistry* / Q000502:physiology; D007700:Kinetics; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry; D013059:Spectrum Analysis, Raman; D054793:Truncated Hemoglobins
chemical_list: D001426:Bacterial Proteins; C102539:GlbN protein, bacteria; D006454:Hemoglobins; D011994:Recombinant Proteins; D054793:Truncated Hemoglobins; D010100:Oxygen