Ligand uptake by M. tuberculosis truncated hemoglobin N is modulated by both two tunnels and retained water molecules

BORON, IGNACIO; BUSTAMANTE, JUAN PABLO; DAVIDGE, KELLY; SINGH, SANDIP; BOWMAN, LESLEY AH; TINAREJO-TREJO, MARIANA; CARBALLAL, SEBASTIAN; RADI, RAFAEL; POOLE, ROBERT K; DIKSHIT, KANAK; ESTRIN, DARÍO; MARTÍ, MARCELO ADRIÁN; BOECHI, LEONARDO

F1000 Research

F1000Research

Año: 2015

2046-1402

Mycobacterium tuberculosis, the causative agent of human tuberculosis, has two proteins belonging to the truncated hemoglobin (trHb) family. Mt-trHbN presents well-defined internal hydrophobic tunnels that allow O2 and ?NO to migrate easily from the solvent to the active site, whereas Mt-trHbO possesses tunnels that are partially blocked by a few bulky residues, particularly a tryptophan at position G8. Differential ligand migration rates allow Mt-trHbN to detoxify ?NO, a crucial step for pathogen survival once under attack by the immune system, much more efficiently than Mt-trHbO. In order to investigate the differences between these proteins, we performed experimental kinetic measurements, ?NO decomposition, as well as molecular dynamics simulations of the wild type Mt-trHbN and two mutants, VG8F and VG8W. These mutations introduce modifications in both tunnel topologies and affect the incoming ligand capacity to displace retained water molecules at the active site. We found that a single mutation allows Mt-trHbN to acquire ligand migration rates comparable to those observed for Mt-trHbO, confirming that ligand migration is regulated by the internal tunnel architecture as well as by water molecules stabilized in the active site.