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Biomedical subjects

L A Segel

Publications and source records attributed to L A Segel.

10 recordsLinked to original sources

Plasmid copy number control: a case study of the quasi-steady-state assumption.

Perelson & Brendel (1989, J. molec. Biol. 208, 245-255) have proposed kinetic models for the control of plasmid copy number, based on experiments by J. Tomizawa and his associates. The quasi-steady-state assumptions (QSSA) made in the analysis of these models are justified in the present paper, thereby providing an example of how QSSA can provide a powerful and reliable tool in the analysis of biological kinetics.

Models, Genetic

Local probes and heterogeneous catalysis: a case study of a mitochondria-luciferase-hexokinase coupled system.

Biological systems are characterized by a high degree of structural organization. In the intracellular context, this introduces physical constraints which are not considered in the standard biochemical analysis of isolated systems, aimed towards mechanistic studies. A major challenge in cellular biology is thus to integrate the structural and mechanistic information and reach an adequate representation of the modes of operation in situ. We present an approach to this problem which takes advantage of a localized probe to study heterogeneous coupled system, as minimal models for cellular operation. The system consists of ATP production at the surface of mitochondria, and ATP consumption in solution by the hexokinase reaction. Soluble or biologically localized firefly luciferase is used to continuously monitor ATP concentration either in the bulk solution or at the surface of the organelle, respectively. The general system of a surface source and a bulk sink is mathematically modeled, and an analytic steady-state solution for local and bulk ATP is presented. The results are validated by experiment and differ from the expected behavior of an equivalent homogeneous system in solution. The model is further adapted to evaluate the effect of mixing. In addition, two limiting cases of heterogeneous distribution of hexokinase are analyzed, in which the soluble enzyme adsorbs non-specifically to mitochondria, or binds selectively to the site of ATP appearance on the membrane. The results are discussed in terms of their significance to the analysis of bulk measurements in vitro and their relevance to better description of cellular situations.

Adenosine Triphosphate

A singular perturbation approach to diffusion reaction equations containing a point source, with application to the hemolytic plaque assay.

Many cells secrete factors which diffuse and bind to receptors on neighboring cells. These processes can be described by a nonlinear diffusion equation with a point source and spatially distributed binding reaction. We show via perturbation analysis how approximate solutions can be obtained for such equations when the binding reaction is fast compared to diffusive transport. We base our analysis on an example which is of great practical importance in immunology, the hemolytic plaque technique.

Binding Sites

Unified mechanism for relay and oscillation of cyclic AMP in Dictyostelium discoideum.

A modified version of an allosteric model for adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] previously analyzed for sustained oscillations of adenosine 3':5'-cyclic monophosphate (cAMP) in Dictyostelium discoideum [Goldbeter, A. (1975) Nature 253, 540-542] is examined to see whether it can account for the relay of cAMP pulses. Oscillations occur around a nonequilibrium, unstable stationary state when system parameters are in a certain domain. It is found that relay can occur outside this domain, in a restricted set of parameter values for which the solution ultimately tends to a stable steady state. A suprathreshold level of extracellular cAMP is needed to elicit relay which consists in a pulsatory synthesis of intracellular cAMP. Theoretical predictions are compared with the results of experiments on cAMP relay and oscillation in aggregation-competent cells of D. discoideum. The model suggests an explanation for the emergence of aggregation centers and for a sequence of developmental events observed in interphase amoebae.

Adenylyl Cyclases

A simple quantitative assay for bacterial motility.

It is argued that the average motility of bacterial populations should be identified with a diffusivity parameter. A simple capillary assay for quantifying this parameter is described, and some results obtained by using this procedure are presented. It is concluded that the assay combine speed and simplicity of operation with sufficient accuracy to make it a valuable tool in the assessment of motility.

Bacterial Physiological Phenomena

Computer evidence concerning the chemotactic signal in Dictyostelium discoideum.

Observed pulsatile aggregation of cellular slime mould amoebae is simulated on a computer. One spatial dimension is considered. In the simulation, attractant is rapidly secreted by the cells, after a delay period, when a superthreshold attractant concentration is sensed. Cells are refractory to further signals after secretion. Once secreted, the attractant diffuses and is hydrolysed. Movement results if a cell's extending pseudopods sense a supercritical increase of attractant; if increases are sensed on both sides, a sufficiently large difference can also initiate movement. The movement continues for a period independently of further signals, but then can be reversed by a attractant increase (at the back of the cell) that surpasses a high threshold. After 100s, motion stops and the threshold for movement reverts to normal. With the above rules, and with parameter values taken, as far as possible, from the literature, the simulation provides the observed pattern of aggration. Outward moving waves of attractant and organized inward pulsatile 'steps' of cell movement surround a cell that sevretes autonomously every few minutes. Other rules fail to give this picture, or give it only for a relatively narrow range of parameter values. It appears that of the various possible signals for chemotaxos, the most likely to be used by the amoebae is a temporal increase of attractant as sensed by extending pseudopods. Nonetheless, we connot rule out the 'classical' hypothesis that cells directly sense concentration differences.

Cell Aggregation