Gamma-secretase inhibition

Development of Alzheimer's disease (AD) pathology appears to be causally related to age-dependent changes in the metabolism of the amyloid-beta peptide (A beta), leading to its enhanced aggregation and deposition. gamma-Secretase is a crucial enzyme for the generation of A beta from the amyloid-beta precursor protein and thus represents a valid potential therapeutic target for the treatment or prevention of AD. Enzyme activity has been shown to be dependent on the expression of presenilins and the identification of inhibitors containing transition-state analogue mimics, together with mutagenesis and knockout studies, confirms that presenilins may provide at least a component of the catalytic site for this putative aspartyl protease. Considerable effort has been expended to identify compounds which specifically reduce gamma-secretase activity in the central nervous system, and those with the appropriate properties are being utilized in on-going proof-of-concept studies in animals and humans, to determine the extent and duration of gamma-secretase inhibition required to elicit therapeutic benefits. gamma-Secretase-mediated substrate cleavage appears to fall into the category of 'regulated intramembrane proteolysis'. By virtue of its mechanistic similarities, the effects of gamma-secretase inhibitors on proteolysis and signalling through other substrates, such as Notch, has to be determined carefully, since this is likely to impact on the clinically safe dose of these compounds.     

Perisomatic inhibition

Recent evidence supports the hypothesis of a functional dichotomy of perisomatic inhibition in the cerebral cortex: the parvalbumin- and cholecystokinin-containing basket cells that are specialized to control rhythm (as a clockwork) and "mood" (as a fine-tuning device), respectively, of network oscillations. Pathology extends this conclusion further, as the former is implicated in epilepsy and the latter in anxiety. The well-balanced cooperation of the two inhibitory systems is required for the normal network operations underlying the cognitive functions of the cerebral cortex.     

Keeping inhibition timely

GABAergic interneurons play a key role in orchestrating cortical network oscillations. In this issue of Neuron, two studies (Bacci and Huguenard and Vida et al.) identify how networks of fast-spiking interneurons can enhance the regularity, precision, and robustness of their own rhythmicity via individual and collective self-innervation.     

Contact inhibition revisited

Contact inhibition of cell movement was originally defined in the 1950s as a way of interpreting studies that were ethological and statistical in nature. Research done in succeeding decades provided a more detailed study of the initial contact and its consequences for the cell. The behavior called contact inhibition is characterized by the cessation of ruffling and forward movement in the lamellipodium of the cell making the contact. A new ruffling membrane then arises elsewhere on the cell perimeter. A comparison between the contact behavior described in the early literature and that of the nerve growth cone, described recently by Steketee and Tosney, suggests that filopodia mediate the sensing function in both cases. Since transformed cells have fewer filopodia than normal cells, the contact behavior may decline in direct response to the degraded function of filopodia. This new “filopodia focal signal transduction” hypothesis of contact inhibition elevates the filopodia sensing function and the cessation of lamellipodial advance to the highest importance as phenomena underlying the altered behavior of cancer cells. J. Cell. Physiol. 220: 574–575, 2009. © 2009 Wiley‐Liss, Inc.     

Diagnosis of enzyme inhibition based on the degree of inhibition

In this work, a method for the diagnosis of kinetic inhibition, based on the dependence of the degree of inhibition (epsilon(i)) on the inhibitor concentration [I] and on the substrate concentration [S], is presented. Because the degree of inhibition is a ratio between rates, kinetic data are normalized by the introduction of an internal control-the rate of the uninhibited reaction. Therefore, the error associated with the kinetic measurements decreases and less experimental measurements are necessary to achieve the diagnosis. The process described, which uses graphical and/or non-linear fitting procedures, allows distinguishing between 20 different kinds of inhibition, including not only linear and hyperbolic, but also parabolic and rational 2,2 inhibitions. Rational 2,2 indicates a new type of inhibition corresponding to an incomplete parabolic inhibition, i.e. mechanistically it corresponds to an inhibitor that binds to two inhibition sites producing enzymatic complexes that are still active. In spite of its comprehensiveness, the diagnosis process is greatly facilitated by the division of the diagnosis of the inhibition in a step-by-step procedure, where only two rival models are evaluated in each step. In the non-linear fittings, the choice between rival models uses a test based on information statistics theory, the Akaike information criterion test, in order to penalize complex models that tend to be favoured in fittings. Finally, equations that allow the determination of inhibition kinetic constants were also deduced. The formalism presented was tested by examining inhibition of acid phosphatase by phosphate (a linear competitive inhibitor).     

On shunting inhibition

The interaction between excitation and inhibition is analyzed for nerve cylinders when reversal potentials for synaptic action are included. Both impulsive and sustained conductance changes are employed to model synaptic action.Exact results, in terms of Green's functions are obtained for the solutions of the cable equation with reversal potentials when there are impulsive conductance changes. The amplification factor for an inhibitory input due to a prior excitatory input is found exactly. In the case of an infinite cylinder, the dependence of this factor on the spatial separation of the excitatory and inhibitory synapses is one plus a Gaussian density function. Similar results aply when excitation follows inhibition. There is shunting inhibition even for impulsive conductance changes in the cable, but it is very different from that for sustained conductance changes. The interaction of excitation and inhibition is also studied in the full cable equation with reversal potentials and sustained conductance changes. An exact result is obtained for the potential in response to simultaneous excitation and inhibition at the same space point in an infinite cable. The effects of timing and spatial separation of inputs is analyzed in a finite nerve cylinder by numerically integrating the cable equation by the Crank-Nicolson method. Shunting inhibition is found to be most effective, for the chosen parameter values, when inhibition quickly foolows excitation. The EPSP amplitude at the soma is found to be roughly proportional to the distance from the soma to the site of inhibition when the excitation is at the center of the nerve cylinder.Dedicated to Jane Pauley     

SHARPINing integrin inhibition

The activity state of integrins is crucial for cell adhesion, migration and differentiation, and is regulated predominantly by protein interactions of the integrin β cytoplasmic domain. SHARPIN is now shown to negatively regulate integrin activation by binding the α-integrin subunit and interfering with the association of the β cytodomain with activating proteins.     

Low oxygen inhibition of photosynthesis is caused by inhibition of starch synthesis

Photosynthesis of C₃ plants is occasionally inhibited upon switching from normal to low partial pressure of O₂. Leaves of Solanum tuberosum exhibited this effect reproducibly under saturating light and 700 microbars of CO₂. We determined the partitioning of recent photosynthate between starch and sucrose and measured the concentration of hexose monophosphates in the stroma and cytosol after nonaqueous fractionation. The reduction in the rate of photosynthesis upon switching to low partial pressure of O₂ was caused by reduced starch synthesis. The concentration of hexose monophosphates in the stroma fell and the glucose 6-phosphate to fructose 6-phosphate to fructose 6-phosphate ratio fell from 2.7 to 1.3, indicating an inhibition of phosphoglucoisomerase as described by K-J Dietz ([1985] Biochim Biophys Acta 839: 240-248). The concentration of hexose monophosphates in the cytosol increased, ruling out a sucrose synthesis limitation by reduced transport from the chloroplast as the explanation for low O₂ inhibition of photosynthesis.     

Specific inhibition of macrophage migration inhibition factor by fucosylated glycolipid RM.

The effects of glycolipids on the interaction of the MIF (migration inhibition factor) with rat macrophages were examined using a migration inhibition assay system. MIF activity was specifically blocked by fucosylated Glycolipid RM [Gal alpha 1-3Gal(2-1 alpha Fuc) beta 1-3GalNAc beta 1-3Gal beta 1-4Glc beta 1-1ceramide, (1978) J. Biochem. 83, 85-90], but not by Cytolipin R, hematoside, or blood group B active glycolipid [Gal alpha 1-3Gal(2-1 alpha Fuc) beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1ceramide]. Inhibition of MIF activity was proportional to the concentration of Glycolipid RM. These findings suggest that Glycolipid RM acts as a receptor for MIF.     

Antigen-mediated macrophage adherence inhibition

Antigen-mediated macrophage adherence inhibition (MAI) was studied in inbred rats immunized with various transplantation, tumour-specific and protein antigens. A macrophage-rich suspension of peritoneal cells (PC) was obtained from the peritoneal cavity of immunized and control animals by washing. The adherence to glass of PC was specifically inhibited by the addition of the antigens used for sensitization of PC donors or by related (cross-reacting) antigens but not with unrelated antigens. The MAI seems to be due to the direct interaction of the respective antigen with a corresponding PC receptor and not due to the humoral factor released from immune lymphocytes of PC population upon contact with the specific antigen.     

Lithium inhibition of phosphofructokinase

The mode of action of lithium in prophylaxis of recurrent affective disorder is unknown although it has been suggested that lithium might compete with magnesium in magnesium dependent processes. We have previously shown pyruvate kinase to be inhibited by lithium and the present study demonstrates a small inhibition by lithium of phosphofructokinase that is also a major regulatory step in glycolysis. Inhibition of PFK was competitive with respect to ATP and magnesium and noncompetitive with respect to potassium and fructose-6-phosphate. Inhibition was enhanced at reduced concentrations of magnesium.