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29 May 2013

978 3 52731 957 2
Wiley VCH
GBP 141.00

Enzyme Kinetics: Principles and Methods

Hans Bisswanger

Enzyme kinetics is an important, but often neglected, subject area for anyone interested in biochemistry, chemical biology, pharmacology, physiology and related sciences. On opening this textbook, the reader is immediately struck by the comprehensive treatment of the subject and the careful development of arguments. The book opens with a useful section defining the various terms that the reader will come across, and is followed by a short introduction and definitions including the differences between thermodynamic and kinetic processes. The latter assumes that the reader has a basic grasp of the subject, although many of the concepts are expanded upon in later chapters.

The main text is arranged into three chapters, equilibria, enzyme kinetics and methods, each with a useful and extensive list of references at the end. The chapter on equilibria covers a large amount of physical (bio)chemistry including the mathematical treatment of diffusion, and the various ligand-binding situations starting from the simplest type. The Michaelis–Menten equation and various methods for analysing data are discussed at this point. More complex situations such as co-operativity and the concerted and sequential models used to explain such behaviour are then discussed, before illustrating these principles using a number of classical examples.

The chapter on enzyme kinetics opens with a discussion of chemical kinetics, covering zero-, first- and second-order situations. The Michaelis–Menten equation is then derived and a number of different methods of determining kinetic parameters are discussed, including the use of the integrated rate equation. More complex behaviour is then covered, including the treatment of reversible enzyme reactions and product inhibition, before moving on to inhibitors. Unusually, this starts with irreversible inhibitors linking the discussion to chemical kinetics and equilibrium processes before moving on to suicide substrates and transition-state analogues. Regrettably, the chemical diagrams used to explain these phenomena contain several errors. Reversible inhibitors are then covered, starting with simple linear inhibitors and moving on to more complex behaviour such as partial inhibition. This is followed by a discussion of multiple substrate enzymes, the various kinetic mechanisms and King–Altmann analysis. The chapter then covers a large amount of material not always contained in similar textbooks, including the treatment of pH effects and protein stability, temperature-dependence effects, isotope exchange and kinetic isotope effects. Particularly nice is the inclusion of a section on special situations including ribozymes, enzymes which use polymer substrates, immobilized enzymes, active transport across membranes and enzyme reactions at membrane surfaces. The chapter closes with a succinct discussion of statistical considerations when analysing kinetic data.

The final chapter deals with experimental methods for determining equilibrium constants and kinetic rates. It starts with the classical physical biochemistry methods for measuring equilibrium binding, including dialysis, ultrafiltration, gel filtration and ultracentrifugation. The chapter then considers more recent techniques, including surface plasmon resonance (SPR), electrochemical methods (the oxygen electrode) and calorimetry. The treatment of UV–visible spectroscopy and fluorescence is particularly thorough, but, unfortunately, the associated chemical diagrams contain several errors. Other spectroscopic methods are then covered, including circular dichroism (CD), infrared spectroscopy, Raman spectroscopy and electron resonance spectroscopy. However, nuclear magnetic resonance (NMR) methods are conspicuous by their absence. The chapter concludes with a discussion of the measurement of fast reactions, using stop flow, relaxation methods, etc. Some of the methods discussed, e.g. electric field method, will have little utility for the study of biological systems, but are included because they are related to biologically relevant methods.

The textbook draws heavily upon physical chemistry and physical biochemistry in its discussion, and the material develops in a logical and progressive manner. Overall, this is an excellent and thorough treatment of a difficult subject. As well as appealing to biochemists, there are also elements of interest to those studying pharmacology and the pharmacokinetic distribution of drugs. The mathematical nature and advanced coverage means that it is most suitable for researchers who already have a strong grounding in the subject. I will certainly be consulting it during my research.


Matthew D. Lloyd (University of Bath)



 
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