Hjem

Preface xi 1 An Introduction to protein structure and function 1 A brief and very selective historical perspective 1 The biological diversity of proteins 5 Proteins and the sequencing of the human and other genomes 9 Why study proteins? 9 2 Amino acids: the building blocks of proteins 13 The 20 amino acids found in proteins 13 The acid-base properties of amino acids 14 Stereochemical representations of amino acids 15 Peptide bonds 16 The chemical and physical properties of amino acids 23 Detection, identification and quantification of amino acids and proteins 32 Stereoisomerism 34 Non-standard amino acids 35 Summary 36 Problems 37 3 The three-dimensional structure of proteins 39 Primary structure or sequence 39 Secondary structure 39 Tertiary structure 50 Quaternary structure 62 The globin family and the role of quaternary structure in modulating activity 66 Immunoglobulins 74 Cyclic proteins 81 Summary 81 Problems 83 4 The structure and function of fibrous proteins 85 The amino acid composition and organization of fibrous proteins 85 Keratins 86 Fibroin 92 Collagen 92 Summary 102 Problems 103 5 The structure and function of membrane proteins 105 The molecular organization of membranes 105 Membrane protein topology and function seen through organization of the erythrocyte membrane 110 Bacteriorhodopsin and the discovery of seven transmembrane helices 114 The structure of the bacterial reaction centre 123 Oxygenic photosynthesis 126 Photosystem I 126 Membrane proteins based on transmembrane β barrels 128 Respiratory complexes 132 Complex III, the ubiquinol-cytochrome c oxidoreductase 132 Complex IV or cytochrome oxidase 138 The structure of ATP synthetase 144 ATPase family 152 Summary 156 Problems 159 6 The diversity of proteins 161 Prebiotic synthesis and the origins of proteins 161 Evolutionary divergence of organisms and its relationship to protein structure and function 163 Protein sequence analysis 165 Protein databases 180 Gene fusion and duplication 181 Secondary structure prediction 181 Genomics and proteomics 183 Summary 187 Problems 187 7 Enzyme kinetics, structure, function, and catalysis 189 Enzyme nomenclature 191 Enzyme co-factors 192 Chemical kinetics 192 The transition state and the action of enzymes 195 The kinetics of enzyme action 197 Catalytic mechanisms 202 Enzyme structure 209 Lysozyme 209 The serine proteases 212 Triose phosphate isomerase 215 Tyrosyl tRNA synthetase 218 EcoRI restriction endonuclease 221 Enzyme inhibition and regulation 224 Irreversible inhibition of enzyme activity 227 Allosteric regulation 231 Covalent modification 237 Isoenzymes or isozymes 241 Summary 242 Problems 244 8 Protein synthesis, processing and turnover 247 Cell cycle 247 The structure of Cdk and its role in the cell cycle 250 Cdk-cyclin complex regulation 252 DNA replication 253 Transcription 254 Eukaryotic transcription factors: variation on a ''basic'' theme 261 The spliceosome and its role in transcription 265 Translation 266 Transfer RNA (tRNA) 267 The composition of prokaryotic and eukaryotic ribosomes 269 A structural basis for protein synthesis 272 An outline of protein synthesis 273 Antibiotics provide insight into protein synthesis 278 Affinity labelling and RNA ''footprinting'' 279 Structural studies of the ribosome 279 Post-translational modification of proteins 287 Protein sorting or targeting 293 The nuclear pore assembly 302 Protein turnover 303 Apoptosis 310 Summary 310 Problems 312 9 Protein expression, purification and characterization 313 The isolation and characterization of proteins 313 Recombinant DNA technology and protein expression 313 Purification of proteins 318 Centrifugation 320 Solubility and ''salting out'' and ''salting in'' 323 Chromatography 326 Dialysis and ultrafiltration 333 Polyacrylamide gel electrophoresis 333 Mass spectrometry 340 How to purify a protein? 342 Summary 344 Problems 345 10 Physical methods of determining the three-dimensional structure of proteins 347 Introduction 347 The use of electromagnetic radiation 348 X-ray crystallography 349 Nuclear magnetic resonance spectroscopy 360 Cryoelectron microscopy 375 Neutron diffraction 379 Optical spectroscopic techniques 379 Vibrational spectroscopy 387 Raman spectroscopy 389 ESR and ENDOR 390 Summary 392 Problems 393 11 Protein folding in vivo and in vitro 395 Introduction 395 Factors determining the protein fold 395 Factors governing protein stability 403 Folding problem and Levinthal''s paradox 403 Models of protein folding 408 Amide exchange and measurement of protein folding 411 Kinetic barriers to refolding 412 In vivo protein folding 415 Membrane protein folding 422 Protein misfolding and the disease state 426 Summary 435 Problems 437 12 Protein structure and a molecular approach to medicine 439 Introduction 439 Sickle cell anaemia 441 Viruses and their impact on health as seen through structure and function 442 HIV and AIDS 443 The influenza virus 457 p53 and its role in cancer 470 Emphysema and α1-antitrypsin 475 Summary 478 Problems 479 Epilogue 481 Glossary 483 Appendices 491 Bibliography 495 References 499 Index 511