Biophysical Chemistry of Proteins [recurso electrónico] : an Introduction to Laboratory Methods

Undergraduate biochemistry courses cover what proteins do, as enzymes, receptors, hormones, motors or structural components. The much more interesting question is how can proteins achieve all these functions? Presented here is an overview of the methods used in such projects, their possible applications, and their limitations. Focusing on the biophysical chemistry of proteins, the text is accessible to those with a general background in chemistry, physics and mathematics, though a good understanding of protein structure and enzymology is required. The text may be used in courses of protein science, by students embarking on master- or PhD-thesis work in this area or by professionals who need a quick reminder about the essentials of a method. Front Matter....Pages i-xvi Front Matter....Pages 1-1 Microscopy....Pages 3-22 Single Molecule Techniques....Pages 23-24 Preparation of Cells and Tissues for Microscopy....Pages 25-26 Principles of Optical Spectroscopy....Pages 27-31 Photometry....Pages 33-37 Fluorimetry....Pages 39-55 Chemiluminescence....Pages 57-60 Electrophoresis....Pages 61-95 Immunological Methods....Pages 97-122 Isotope Techniques....Pages 123-132 Front Matter....Pages 133-133 Homogenisation and Fractionisation of Cells and Tissues....Pages 135-139 Isolation of Organelles....Pages 141-141 Precipitation Methods....Pages 143-146 Chromatography....Pages 147-162 Membrane Proteins....Pages 165-182 Determination of Protein Concentration....Pages 183-186 Cell Culture....Pages 187-189 Front Matter....Pages 191-192 General Technical Remarks....Pages 193-197 Amine-Reactive Reagents....Pages 199-203 Thiol and Disulphide Reactive Reagents....Pages 205-207 Front Matter....Pages 191-192 Reagents for Other Groups....Pages 209-217 Cross-linkers....Pages 219-221 Detection Methods....Pages 223-227 Spontaneous Reactions in Proteins....Pages 229-233 Front Matter....Pages 235-235 Centrifugation....Pages 237-249 Osmotic Pressure....Pages 251-253 Diffusion....Pages 255-256 Viscosity....Pages 257-259 Non-resonant Interactions with Electromagnetic Waves....Pages 261-267 Front Matter....Pages 269-269 Protein Sequencing....Pages 271-288 Synthesis of Peptides....Pages 289-290 Protein Secondary Structure....Pages 291-302 Structure of Protein–Ligand Complexes....Pages 303-308 3-D Structures....Pages 309-342 Folding and Unfolding of Proteins....Pages 343-352 Front Matter....Pages 253-253 Steady-State Kinetics....Pages 355-366 Leaving the Steady State: Analysis of Progress Curves....Pages 367-367 Reaction Velocities....Pages 369-381 Isotope Effects....Pages 383-385 Isotope Exchange....Pages 387-388 Front Matter....Pages 289-290 General Conditions for Interpretable Results....Pages 391-392 Binding Equations....Pages 393-397 Methods to Measure Binding Equilibria....Pages 399-407 Temperature Effects on Binding Equilibrium and Reaction Rate....Pages 409-413 Front Matter....Pages 415-415 Industrial Enzyme Use....Pages 417-420 Immobilised Enzymes....Pages 421-423 Front Matter....Pages 425-425 Quality Control....Pages 427-434 Testing Whether or Not a Model Fits the Data....Pages 435-438 Front Matter....Pages 439-439 List of Symbols....Pages 441-444 Greek Alphabets....Pages 445-445 Properties of Electrophoretic Buffers....Pages 447-452 Bond Properties....Pages 453-453 Acronyms....Pages 455-463 Back Matter....Pages 465-510 Annotation 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 Provides 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. This title is suitable for Scientists interested in protein research