Structural characterization of metal binding to a cold-adapted frataxin

NOGUERA ME; ROMAN EA; RIGAL RB; COUSIDO-SIAH A; MITSCHLER A; PODJARNY A; SANTOS J

JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY

SPRINGER

Lugar: Berlin; Año: 2015 vol. 20 p. 653 - 664

0949-8257

Life adaptation to low temperatures influences both protein stability and flexibility. Thus, proteins from psychrophilic organisms are excellent models to study relations between these properties. Here we focused on frataxin from Psychromonas ingrahamii (pFXN), an extreme psychrophilic sea ice bacterium which can grow at temperatures as low as -12 °C. This α/β protein is highly conserved and plays a key role in iron homeostasis as an iron chaperone. In contrast to other frataxin homologs, chemical and temperature unfolding experiments showed that the thermodynamic stability of pFXN is strongly modulated by pH: ranging from 5.5±0.9 (pH 6.0) to 0.9±0.3 (pH 8.0) kcal mol-1. This protein was crystallized and its X-ray structure solved at 1.45 Å. Comparison of B-factor profiles between E. coli and P. ingrahamii frataxin variants (51% of identity) suggests that, although both proteins share the same structural features, their flexibility distribution is different. Molecular dynamics simulations showed that protonation of His44 or His67 in pFXN lowers the mobility of regions encompassing residues 20-30 and the C-terminal end, probably through favorable electrostatic interactions with residues Asp27, Glu42 and Glu99. Since the C-terminal end of the protein is critical for the stabilization of the frataxin fold, the predictions presented may be reporting on the microscopic origin of the global stability decrease produced near neutral pH in the psychrophilic variant. We propose that suboptimal electrostatic interactions may have been an evolutionary strategy for the adaptation of frataxin flexibility and function to cold environments.