Henry A. Wallace to John D. Feerick

Letter from Vice President Henry A. Wallace to Dean John D. Feerick, regarding his scholarly article on presidential inability.https://ir.lawnet.fordham.edu/twentyfifth_amendment_correspondence/1009/thumbnail.jp

Strategies for studying permeation at voltage-gated ion channels

Voltage-dependent ion channels are presently thought to consist of several distinct functional regions: (a) activation gates, (b) inactivation gates, and permeation pathways. This chapter focuses on permeation pathways and may spur new ideas about experiments that use site-directed mutagenesis to probe the ion conduction pathway. Some hubris is required to attempt a survey of this field since individual families -- K^+, Na^+, or Ca^(2+) -- have been reviewed in detail (15, 68, 115, 127). My unified treatment is motivated by the structural similarity suggested by recent cDNA sequencing data on this group (see, for instance, 24). There have been many excellent previous treatments of ion channel permeation (6, 15, 34, 35, 51, 53, 68, 73, 74, 115, 127)

New Views of Multi-Ion Channels

Thus, most site-directed mutagenesis data render it untenable to consider that two or more roughly equivalent high affinity sites govern selectivity in multi-ion pores. The papers by Dang and McCleskey and Kiss et al. respond to this challenge by showing that a model with a single high affinity site, flanked by two binding sites of lower affinity close to the pore entrances, can generate much of the classical multi-ion behavior. The sites need not interact, and the two flanking sites could arise from one of several mechanisms: a featureless charged vestibule, a dehydration step, or a specific weak binding site. The multi-ion pore remains a cornerstone of permeation theory, but the new theory features only a single high affinity site and no mutual repulsion. The high flux rate occurs because ions pause at the flanking sites and reequilibrate thermally, gaining enough energy to move over the next barrier

Evaluating the operations capability of Freedom's Data Management System

Three areas of Data Management System (DMS) performance are examined: raw processor speed, the subjective speed of the Lynx OS X-Window system, and the operational capacity of the Runtime Object Database (RODB). It is concluded that the proposed processor will operate at its specified rate of speed and that the X-Window system operates within users' subjective needs. It is also concluded that the RODB cannot provide the required level of service, even with a two-order of magnitude (100 fold) improvement in speed

OPPORTUNITIES FOR EFFECTIVE EXTENSION POLICY EDUCATION

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An Intermediate State of the {gamma}-Aminobutyric Acid Transporter GAT1 Revealed by Simultaneous Voltage Clamp and Fluorescence

The rat {gamma}-aminobutyric acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly altering GAT1 function. Voltage-jump relaxation analysis showed that fluorescence increased slightly and monotonically with hyperpolarization; the fluorescence at -140 mV was ~0.8% greater than at +60 mV. The time course of the fluorescence relaxations was mostly described by a single exponential with voltage-dependent but history-independent time constants ranging from ~20 ms at +60 mV to ~150 ms at -140 mV. The fluorescence did not saturate at the most negative potentials tested, and the midpoint of the fluorescence–voltage relation was at least 50 mV more negative than the midpoint of the charge–voltage relation previously identified with Na+ binding to GAT1. The presence of {gamma}-aminobutyric acid did not noticeably affect the fluorescence waveforms. The fluorescence signal depended on Na+ concentration with a Hill coefficient approaching 2. Increasing Cl- concentration modestly increased and accelerated the fluorescence relaxations for hyperpolarizing jumps. The fluorescence change was blocked by the GAT1 inhibitor, NO-711. For the W68L mutant of GAT1, the fluorescence relaxations occurred only during jumps to high positive potentials, in agreement with previous suggestions that this mutant is trapped in one conformational state except at these potentials. These observations suggest that the fluorescence signals monitor a novel state of GAT1, intermediate between the E*out and Eout states of Hilgemann, D.W., and C.-C. Lu (1999. J. Gen. Physiol. 114:459–476). Therefore, the study provides verification that conformational changes occur during GAT1 function

OPPORTUNITIES FOR PUBLIC POLICY EDUCATION IN THE EXTENSION INITIATIVES

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