Ebook: Biophysical Chemistry of Proteins: 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.

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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.

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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.
Content:
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

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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.
Content:
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
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