Main description:

Proteins constitute the working-class molecules of the cell. Hence, understanding the way they act is a prerequisite for understanding how a cell functions and how life evolves. Aspects such as the protein-ligand relationship, recognition, protein evolution by point mutation, enzyme-substrate interactions, behaviour of an enzyme in a living cell, control and dynamics of enzyme networks as well as the physico-chemical background of enzyme actions and multi-enzyme complexes are comprehensively treated in this volume.

Contents:

Dynamics of Enzyme Reactions and Metabolic Networks in Living Cells. A Physico-Chemical Approach.- 1. Introduction.- 2. Flows and forces, diffusion, partition of mobile ions by charged matrices.- 3. Compartmentalization of enzyme reactions and the energy metabolism of the cell.- 4. Coupling between reactant diffusion and bound enzyme reaction rate.- 5. Electric partitioning of ions and reaction rate of bound enzyme systems.- 5.1. Electrostatic co-operativity of a bound enzyme system.- 5.2. Spatial organization of fixed charges and enzyme molecules as a source of co-operativity.- 6. An example of enzyme behaviour in organized biological systems: the dynamics of enzymes bound to plant cell walls.- 7. Control and dynamics of enzyme networks.- 7.1. Control of a linear metabolic network.- 7.2. Dynamic organization of a metabolic cycle in homogeneous phase.- 7.3. Dynamic organization of a metabolic cycle at the surface of a charged membrane.- 8. Control of multi-enzyme complexes.- 8.1. Generalized microscopic reversibility and multi-enzyme complexes.- 8.2. The stoichiometry of polypeptide chains in multi-enzyme complexes.- 8.3. The principles of structural kinetics of oligomeric enzymes and of multi-enzyme complexes.- 8.4. Thermodynamics of the alteration of the kinetic behaviour of an oligomeric enzymes within a multi-enzyme complex.- 9. General Conclusions.- References.- Microbial and Genetic Approaches to the Study of Structure-Function Relationships of Proteins.- 1. Introduction.- 2. Protein-ligand and protein-substrate interactions.- 2.1. The recognition problem.- 2.2. Conformational selection by ligand interaction.- 2.3. Protein dynamics and simulation.- 3. Protein evolution by point mutation.- 3.1. The suppressor approach: position-specific design of the substitution matrix.- 3.2. Interdependence of residues located in the hydrophobic core.- 3.3. Essential residues revisited.- 3.4. What is a neutral position?.- 4. Protein evolution by rearrangements of combinatorial domains.- 4.1. Acquiring and integrating new domain(s).- 4.2. Protein secretion in Gram-negative bacteria.- 4.2.1. Domain recruitment as a strategy for acquiring a new function.- 4.2.2. Domain-domain interaction as a strategy for self and non-self recognition.- 5. Conclusion and perspectives.- References.- Recent Progress in Studies of Enzymatic Systems in Living Cells.- 1. Introduction: Why study the behaviour of enzymes in single living cells?.- 1.1. Some words about fluorescence.- 1.2. Practical consequences.- 2. Tools available for studying the behavior of enzymes in single living cells.- 2.1. Instrumentation.- 2.1.1. Fluorescence anisotropy of cells suspension.- 2.1.2. Flow cytometry.- 2.1.3. Microfluorometry (spectral mode).- 2.1.4. Microfluorometry (topographic mode).- 2.1.5. Videomicrofluorometry.- 2.1.6. Life time measurements (decay of intensity and phase fluorometry).- 2.2. Progress in software and modelization.- 2.2.1. Spectral resolution.- 2.2.2. Correlation with biochemical conventional data.- 3. What next?.- 4. General conclusion.- References.