Potassium Transport and the Relationship Between Intracellular Potassium Concentration and Amino Acid Uptake by Cells of a Marine Pseudomonad

Transport of K(+) by K(+)-depleted cells of marine pseudomonad B-16 (ATCC 19855) exhibited saturation kinetics. Rb(+) inhibited both K(+) transport and the K(+)-dependent transport of alpha-aminoisobutyric acid (AIB) into K(+)-depleted cells of the organism in proportion to the concentration of Rb(+) in the suspending medium. Inhibition of the K(+)-dependent uptake of AIB into K(+)-depleted cells by Rb(+) could be overcome by increasing the concentration of K(+) in the medium. When AIB and K(+) were added simultaneously to a suspension of K(+)-depleted cells, the uptake of K(+) occurred immediately and rapidly, whereas the accumulation of AIB occurred only after a lag. The initial uptake rate of AIB was directly proportional to the intracellular K(+) concentration. The intracellular concentration of K(+) and AIB at their steady-state levels increased to a maximum as the Na(+) concentration in the suspending medium was increased. At Na(+) concentrations between 0.2 and 0.3 M, the molar ratio of K(+) to AIB at their intracellular steady-state concentrations was constant at 1.6. At external Na(+) concentrations less than 0.2 M, the cells maintained a relatively higher K(+) intracellular steady-state level than AIB.     

Biochemical Localization of Alkaline Phosphatase in the Cell Wall of a Marine Pseudomonad

The various layers of the cell envelope of marine pseudomonad B-16 (ATCC 19855) have been separated from the cells and assayed directly for alkaline phosphatase activity under conditions established previously to be optimum for maintenance of the activity of the enzyme. Under conditions known to lead to the release of the contents of the periplasmic space from the cells, over 90% of the alkaline phosphatase was released into the medium. Neither the loosely bound outer layer nor the outer double-track layer (cell wall membrane) showed significant activity. A small amount of the alkaline phosphatase activity of the cells remained associated with the mureinoplasts when the outer layers of the cell wall were removed. Upon treatment of the mureinoplasts with lysozyme, some alkaline phosphatase was released into the medium and some remained with the protoplasts formed. Cells washed and suspended in 0.5 M NaCl were lysed by treatment with 2% toluene, and 95% of the alkaline phosphatase in the cells was released into the medium. Cells washed and suspended in complete salts solution (0.3 M NaCl, 0.05 M MgSO(4), and 0.01 M KCl) or 0.05 M MgSO(4) appeared intact after treatment with toluene but lost 50 and 10%, respectively, of their alkaline phosphatase. The results suggest that the presence of Mg(2+) in the cell wall is necessary to prevent disruption of the cells by toluene and may also be required to prevent the release of alkaline phosphatase by toluene when disruption of the cells by toluene does not take place.     

Mechanical properties of muscle spindles in Xenopus laevis

Mechanical properties of isolated living muscle spindles from Xenopus laevis were examined in order to determine their role in sensory transduction. The reticular zone of the intrafusal muscle fibers was identified microscopically by: (1) its position beneath the sensory endings, (2) its length, 50–100 μm, (3) its extension during intrafusal muscle contraction, and (4) its coarse striations with a period of about 1.5 times the normal sarcomere length. The reticular zone in the passive muscle spindle did not extend until the spindle was stretched to about 1.05–1.1 its maximal length in the animal (L _m ). Evidence was obtained that the absence of extension of the reticular zone at normal muscle lengths was due to the presence of the spindle capsule which acted as a stiff element in parallel with the sensory region. At those lengths at which the reticular zone did extend (> L _m ), no rate — sensitive mechanical properties were detected in response to step and ramp extensions. The sensory discharge of the spindle showed no dynamic transient in response to ramp extensions if the reticular zone were not extended. During extension of the reticular zone a dynamic sensory transient appeared. It is concluded that current notions on the mechanical origin of the rate — sensitive properties of the sensory discharge of the muscle spindle do not apply to Xenopus laevis. In addition, it is not likely that the passive spindle in this animal is a sensitive stretch receptor.