Full-scale model of glycolysis in Saccharomyces cerevisiae.

We present a powerful, general method of fitting a model of a biochemical pathway to experimental substrate concentrations and dynamical properties measured at a stationary state, when the mechanism is largely known but kinetic parameters are lacking. Rate constants and maximum velocities are calculated from the experimental data by simple algebra without integration of kinetic equations. Using this direct approach, we fit a comprehensive model of glycolysis and glycolytic oscillations in intact yeast cells to data measured on a suspension of living cells of Saccharomyces cerevisiae near a Hopf bifurcation, and to a large set of stationary concentrations and other data estimated from comparable batch experiments. The resulting model agrees with almost all experimentally known stationary concentrations and metabolic fluxes, with the frequency of oscillation and with the majority of other experimentally known kinetic and dynamical variables. The functional forms of the rate equations have not been optimized.

Synchronization of glycolytic oscillations in a yeast cell population.

The mechanism of active phase synchronization in a suspension of oscillatory yeast cells has remained a puzzle for almost half a century. The difficulty of the problem stems from the fact that the synchronization phenomenon involves the entire metabolic network of glycolysis and fermentation, and consequently it cannot be addressed at the level of a single enzyme or a single chemical species. In this paper it is shown how this system in a CSTR (continuous flow stirred tank reactor) can be modelled quantitatively as a population of Stuart-Landau oscillators interacting by exchange of metabolites through the extracellular medium, thus reducing the complexity of the problem without sacrificing the biochemical realism. The parameters of the model can be derived by a systematic expansion from any full-scale model of the yeast cell kinetics with a supercritical Hopf bifurcation. Some parameter values can also be obtained directly from analysis of perturbation experiments. In the mean-field limit, equations for the study of populations having a distribution of frequencies are used to simulate the effect of the inherent variations between cells.

Sustained oscillations in living cells.

Glycolytic oscillations in yeast have been studied for many years simply by adding a glucose pulse to a suspension of cells and measuring the resulting transient oscillations of NADH. Here we show, using a suspension of yeast cells, that living cells can be kept in a well defined oscillating state indefinitely when starved cells, glucose and cyanide are pumped into a cuvette with outflow of surplus liquid. Our results show that the transitions between stationary and oscillatory behaviour are uniquely described mathematically by the Hopf bifurcation. This result characterizes the dynamical properties close to the transition point. Our perturbation experiments show that the cells remain strongly coupled very close to the transition. Therefore, the transition takes place in each of the cells and is not a desynchronization phenomenon. With these two observations, a study of the kinetic details of glycolysis, as it actually takes place in a living cell, is possible using experiments designed in the framework of nonlinear dynamics. Acetaldehyde is known to synchronize the oscillations. Our results show that glucose is another messenger substance, as long as the glucose transporter is not saturated.

Systematic derivation of amplitude equations and normal forms for dynamical systems.

We present a systematic approach to deriving normal forms and related amplitude equations for flows and discrete dynamics on the center manifold of a dynamical system at local bifurcations and unfoldings of these. We derive a general, explicit recurrence relation that completely determines the amplitude equation and the associated transformation from amplitudes to physical space. At any order, the relation provides explicit expressions for all the nonvanishing coefficients of the amplitude equation together with straightforward linear equations for the coefficients of the transformation. The recurrence relation therefore provides all the machinery needed to solve a given physical problem in physical terms through an amplitude equation. The new result applies to any local bifurcation of a flow or map for which all the critical eigenvalues are semisimple (i.e., have Riesz index unity). The method is an efficient and rigorous alternative to more intuitive approaches in terms of multiple time scales. We illustrate the use of the method by deriving amplitude equations and associated transformations for the most common simple bifurcations in flows and iterated maps. The results are expressed in tables in a form that can be immediately applied to specific problems. (c) 1998 American Institute of Physics.

Sustained oscillations in glycolysis: an experimental and theoretical study of chaotic and complex periodic behavior and of quenching of simple oscillations.

We report sustained oscillations in glycolysis conducted in an open system (a continuous-flow, stirred tank reactor; CSTR) with inflow of yeast extract as well as glucose. Depending on the operating conditions, we observe simple or complex periodic oscillations or chaos. We report the response of the system to instantaneous additions of small amounts of several substrates as functions of the amount added and the phase of the addition. We simulate oscillations and perturbations by a kinetic model based on the mechanism of glycolysis in a CSTR. We find that the response to particular perturbations forms an efficient tool for elucidating the mechanism of biochemical oscillations.

Chaos in Glycolysis

Glycolysis occurs in almost every living cell as part of the energy metabolism. It forms a complex dynamical system, and might thus be capable of exhibiting complex phenomena. Simple oscillations have been observed frequently in suspensions of intact cells and in cell extracts, but only as transients. We have obtained sustained simple and complex oscillations in glycolysis of cell-free yeast extract in a flow-reactor. Sustained oscillations enable a powerful, proven method of dynamical system theory to unravel the kinetics and make it possible to observe chaos. Chaos was predicted from models long ago but has not previously been observed experimentally. We report the first experimental observation of unforced chaotic oscillations in glycolysis. Copyright 1997 Academic Press Limited

The Ginzburg-Landau approach to oscillatory media.

Close to a supercritical Hopf bifurcation, oscillatory media may be described, by the complex Ginzburg-Landau equation. The most important spatiotemporal behaviors associated with this dynamics are reviewed here. It is shown, on a few concrete examples, how real chemical oscillators may be described by this equation, and how its coefficients may be obtained from the experimental data. Furthermore, the effect of natural forcings, induced by the experimental realization of chemical oscillators in batch reactors, may also be studied in the framework of complex Ginzburg-Landau equations and its associated phase dynamics. We show, in particular, how such forcings may locally transform oscillatory media into excitable ones and trigger the formation of complex spatiotemporal patterns.