Swimming behavior of the monotrichous bacterium Pseudomonas fluorescens SBW25.

Motility is an important trait for some bacteria living in nature and the analyses of it can provide important information on bacterial ecology. While the swimming behavior of peritrichous bacteria such as Escherichia coli has been extensively studied, the monotrichous bacteria such as the soil inhabiting and plant growth promoting bacterium Pseudmonas fluorescens is not very well characterized. Unlike E. coli that is propelled by a left-handed flagella bundle, P. fluorescens SBW25 swims several times faster by rotating a right-handed flagellum. Its swimming pattern is the most sophisticated known so far: it swims forward (run) and backward (backup); it can swiftly 'turn' the run directions or 'reorient' at run-backup transitions; it can 'flip' the cell body continuously or 'hover' in the milieu without translocation. The bacteria swam in circles near flat surfaces with reduced velocity and increased turn frequency. The viscous drag load due to wall effect potentially accounts for the circular motion and velocity change, but not the turn frequency. The flagellation and swimming behavior of P. fluorescens SBW25 show some similarity to Caulobacter, a fresh-water inhabitant, while the complex swimming pattern might be an adaptation to the geometrically restricted rhizo- and phyllospheres.

Oxidation of multiple methionine residues impairs rapid sodium channel inactivation.

Reactive oxygen species (ROS) readily oxidize the sulfur-containing amino acids cysteine and methionine (Met). The impact of Met oxidation on the fast inactivation of the skeletal muscle sodium channel Na(V)1.4 expressed in mammalian cells was studied by applying the Met-preferring oxidant chloramine-T or by irradiating the ROS-producing dye Lucifer Yellow in the patch pipettes. Both interventions dramatically slowed down inactivation of the sodium channels. Replacement of Met in the Ile-Phe-Met inactivation motif with Leu (M1305L) strongly attenuated the oxidizing effect on inactivation but did not eliminate it completely. Mutagenesis of Met1470 in the putative receptor of the inactivation lid also markedly diminished the oxidation sensitivity of the channel, while that of other conserved Met residues in intracellular linkers connecting the membrane-spanning segments (442, 1139, 1154, 1316, 1469) were of minor importance. The results of mutagenesis, assays of other Na(V) channel isoforms (Na(V)1.2, Na(V)1.5, Na(V)1.7), and the kinetics of the oxidation-induced removal of inactivation collectively indicate that multiple Met residues need to be oxidized to completely impair inactivation. This arrangement using multiple Met residues confers a finely graded oxidative modulation of Na(V) channels and allows organisms to adapt to a variety of oxidative stress conditions, such as ischemic reperfusion.