Biophysical Chemistry of Proteins, 2011 An Introduction to Laboratory Methods

The book is structured in nine sections, each containing several chapters. The volume starts with an overview of analytical techniques and progresses through purification of proteins; protein modification and inactivation; protein size, shape, and structure; enzyme kinetics; protein-ligand interactions; industrial enzymology; and laboratory quality control. The book is targeted at all scientists interested in protein research.

Analytical techniques. Microscopy. Single molecule techniques. Preparation of cells and tissues for microscopy. Principles of optical spectroscopy.Photometry. Fluorimetry. Chemiluminescence.Electrophoresis.Immunological methods.Isotope techniques.- Purification of proteins.Homogenisation and fractionisation of cells and tissues.Isolation of organelles. Precipitation methods.Chromatography. Membrane proteins. Determination of protein concentration.Cell culture.- Protein modification and inactivation. General technical remarks. Amine-reactive reagents. Thiol- and disulphide reactive reagents. Reagents for other groups. Cross-linkers.Detection methods. Spontaneous reactions in proteins.- Protein size and shape. Centrifugation.Osmotic pressure. Diffusion.Viscosity.Non-resonant interactions with electromagnetic waves.- Protein structure. Protein sequencing.Synthesis of peptides. Protein secondary structure. Structure of protein-ligand complexes. 3D-structures. Folding and unfolding of proteins.- Enzyme kinetics. Steady-state kinetics. Leaving the steady state: Analysis of progress curves.Reaction velocities. Isotope effects. Isotope exchange.- Protein-ligand interactions. General conditions for interpretable results. Binding equations.Methods to measure binding equilibria. Temperature effects on binding equilibrium and reaction rate.- Industrial enzymology. Industrial enzyme use.Immobilised enzymes.- Special statistics.Quality control. Testing whether or not a model fits the data.- Appendix. List of symbols.Greek alphabet. Properties of electrophoretic buffers. Bond properties. Acronyms.

Dr. Buxbaum's research interests are enzymology and protein structure/function relationship. In addition, he has been teaching science and medical students in several countries. He is currently working as associate professor of biochemistry at Ross University School of Medicine.

Focuses on the biophysical chemistry of proteins Presents an overview of the methods used in protein research Examines the possible applications and limitations of protein research