Interaction of Tetraethylammonium Ion Derivatives with the Potassium Channels of Giant Axons

A number of compounds related to TEA⁺ (tetraethylammoniumion) were injected into squid axons and their effects on g_K (the potassium conductance) were determined. In most of these ions a quaternary nitrogen is surrounded by three ethyl groups and a fourth group that is very hydrophobic. Several of the ions cause inactivation of g_K, a type of ionic gating that is not normally seen in squid axon; i.e., after depolarization g_K increases and then spontaneously decreases to a small fraction of its peak value even though the depolarization is maintained. Observations on the mechanism of this gating show that (a) QA (quaternary ammonium) ions only enter K⁺ channels that have open activation gates (the normal permeability gates). (b) The activation gates of QA-occluded channels do not close readily. (c) Hyperpolarization helps to clear QA ions from the channels. (d) Raising the external K⁺ concentration also helps to clear QA ions from the channels. Observations (c) and (d) strongly suggest that K⁺ ions traverse the membrane by way of pores, and they cannot be explained by the usual type of carrier model. The data suggest that a K⁺ pore has two distinct parts: a wide inner mouth that can accept a hydrated K⁺ ion or a TEA⁺-like ion, and a narrower portion that can accept a dehydrated or partially dehydrated K⁺ ion, but not TEA⁺.     

Alkaline phosphatase of Drosophila melanogaster. II. Biochemical comparison among four allelic forms

Biochemical analyses of partially purified preparations of APH-4 and -6 (common allelic forms) and APH-2 and -10 (rare allelic forms) of D. melanogaster reveal that the two common forms are similar in all properties investigated except for pH optimum (8.0 vs. 8.5). The common and rare forms share certain properties in common but differ in that the common forms are more stable to heat and more sensitive to inhibition by inorganic phosphate. With respect to such properties as substrate preferences and K _i values for inorganic phosphate, the common forms and APH-2 are similar to one another, whereas APH-10 is distinctly different. All four activities show preference for a phosphoaromatic compound as substrate, with O-phosphotyrosine being the best substrate of biological origin. Transphosphorylation, as related to these allelic forms of APH, is discussed.Paper No. 3892 of the Journal Series of the North Carolina State University Agricultural Experiment Station, Raleigh, North Carolina. This study was supported by Atomic Energy Commission Contract AT-(40-1-)-3980.     

The reversal of phenylarsenoxide inhibition of keto acid oxidation in mitochondrial and bacterial suspensions by lipoic acid and other disulphides

1. Inhibition of pyruvate oxidation in suspensions of Aerobacter aerogenes cells and of isolated mitochondria from rat heart and liver by phenylarsenoxide is prevented by an excess of lipoic acid, whereas inhibition due to certain bivalent cations is not. 2. In both systems inhibition persists when the bacteria and mitochondria are recovered and resuspended in fresh media in the absence of the inhibitor. Persistent inhibition due to preincubation with phenylarsenoxide, but not with the metal ions, is reversed by lipoic acid and by certain other disulphides. 3. 2,3-Dimercaptopropan-1-ol prevents the inhibition of pyruvate oxidation by phenylarsenoxide and by bivalent cations in both mitochondria and bacterial cells. 4. In aerobic suspensions of mitochondria and bacteria disulphides such as lipoic acid are reduced rapidly to dithiols. Reduction is inhibited by Co(2+), Ni(2+), Cd(2+) and Zn(2+), but not by phenylarsenoxide. 5. It is concluded that the inability of lipoic acid to prevent the action of the metal ions on pyruvate oxidation is due to the inhibition of its reduction to the effective dithiol.