The physical and chemical basis of molecular biology:
Gespeichert in:
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| Format: | Buch |
| Sprache: | English |
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[Eastbourne]
Helvetian Press
2010
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| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XXXIX, 642 S. Ill., graph. Darst. |
| ISBN: | 9780956478108 |
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| 084 | |a PHY 821f |2 stub | ||
| 084 | |a BIO 220f |2 stub | ||
| 100 | 1 | |a Creighton, Thomas E. |e Verfasser |4 aut | |
| 245 | 1 | 0 | |a The physical and chemical basis of molecular biology |c Thomas E. Creighton |
| 246 | 1 | 3 | |a Molecular biology |
| 264 | 1 | |a [Eastbourne] |b Helvetian Press |c 2010 | |
| 300 | |a XXXIX, 642 S. |b Ill., graph. Darst. | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 650 | 0 | 7 | |a Molekulare Biophysik |0 (DE-588)4170391-1 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Molekularbiologie |0 (DE-588)4039983-7 |2 gnd |9 rswk-swf |
| 650 | 0 | 7 | |a Biochemie |0 (DE-588)4006777-4 |2 gnd |9 rswk-swf |
| 689 | 0 | 0 | |a Molekularbiologie |0 (DE-588)4039983-7 |D s |
| 689 | 0 | 1 | |a Molekulare Biophysik |0 (DE-588)4170391-1 |D s |
| 689 | 0 | 2 | |a Biochemie |0 (DE-588)4006777-4 |D s |
| 689 | 0 | |C b |5 DE-604 | |
| 856 | 4 | 2 | |m Digitalisierung UB Regensburg |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020534011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 999 | |a oai:aleph.bib-bvb.de:BVB01-020534011 | ||
Datensatz im Suchindex
| _version_ | 1804143220507017216 |
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| adam_text | CONTENTS
Preface
xix
Common Abbreviations
xxii
Glossary wxi
Section I: Fundamentals
1.
Thermodynamics for molecular biology
1
1.1.
Equilibrium constants
1
1.2
Gibbs free energy
3
1.2.
A. Coupled reactions
5
1.2.В.
Linked functions
6
1.3.
Enthalpy
7
1.4.
Entropy
9
1.5.
Heat capacity
11
1.6.
Calorimetry
13
1.6.
A. Isothermal titration calorimetry
14
1.6.В.
Differential scanning calorimetry
16
2.
Noncovalent interactions between atoms and molecules
18
2.1.
Short-range repulsions: defining atomic volume
18
2.I.A.
Molecular surfaces and volumes
20
2.1.В.
Packing density
21
2.2.
Electrostatic forces: simplicity to complexity
22
2.2.
A. Point charges: the simplest interaction
22
2.2.B. Dipoles: charge separation within a molecule
23
1. Dipole
moment
25
2.
Polarizability
26
2.2.
С
Ion pairs and salt bridges
26
2.3.
Van
der Waals
interactions: the advantages of close packing
27
2.4.
Hydrogen bonds: specificity and directionality
29
2.5.
Intramolecular interactions: the importance of entropy
32
2.5.
A. Effective concentrations: an empirical approach to the
entropy problem
33
vi
CONTENTS
2.5.
В.
Multiple
interactions:
entropy and cooperativity
36
2.5.
С.
Cooperativity of multiple interactions: the key to
macromolecule
folding
37
3.
Aqueous solutions
42
3.1.
Liquid water
43
3.1.
A. Liquids: close interactions without order
43
3.1.
B. Water: the importance of hydrogen bonding
45
3.2.
The hydrophobic interaction: avoiding waters phobia
49
3.2.
A. Partition coefficients: measuring preferences for different
environments
50
3.2.B. The hydrophobic interaction in
nonpolar
model systems
51
3.3.
Membranes: hydrophobic bilayers in an aqueous environment
58
3.3.A. Detergents
65
3.4.
Ionization
67
3.4.
A. Measuring the
pH 71
3.4.B. Buffers
72
1.
Phosphate buffers
74
2.
Tris
buffer
75
3.
Membrane-impermeable Good buffers
76
4.
Volatile buffers
76
3.5.
Salts and ions
77
3.6.
Electrostatic interactions in water: Debye and Hiickel
79
3.6.
A. Poisson
and Boltzmann
80
3.7.
Solubilities in water
80
3.7.
A. Salting in, salting out
81
3.8. Hofmeister
series
83
3.9. Hydration
of macromolecules
84
3.9.
A. Preferential hydration versus preferential binding
85
3.9.
B. Transfer free energy
88
3.10.
Chemical potential
88
3.11.
Compressibility: the effects of high pressure
89
4.
Kinetics: a brief review
91
4.1.
Single reactions
91
4.I.A.
First-order kinetics
93
1.
Half-time
95
2.
Relaxation time
96
3.
Reversible reactions
96
4.1.В.
Second-order kinetics
97
4.
l.C. Zero-order kinetics
99
4.I.D.
Transition state
99
4.
I.E. Free-energy relationships
104
CONTENTS
vii
4.2.
Multi-step reactions and intermediates
105
4.2.
A. Rate-determining step
107
4.2.B. Steady-state kinetics
110
4.3.
Measuring rapid reactions
110
4.3.A. Rapid mixing techniques 111
4.3.
B. Relaxation techniques
113
5.
Isotopes and radioactivity
115
5.1.
Isotopes
115
5.2.
Radioactive decay
116
5.2.A. Alpha particles
118
5.2.B. Beta particles
119
5.2.C. Positrons
120
5.2.D. Gamma rays
120
5.3.
Kinetics of radioactive decay
120
5.3.A. Units
121
5.4.
Measurement of radioactivity
121
5.4.
A. Radiation counters
121
1.
Ionization monitor
122
2.
Scintillation counters
122
3.
Cerenkov radiation
123
5.4.B. Autoradiography
123
1.
Film-less autoradiography
124
2.
Fluorography
125
5.5. Radioisotopes
commonly used in molecular biology
125
5.5.
A. Hydrogen isotopes
125
1.
Tritium
125
2.
Deuterium
127
5.5.B. Carbon isotopes
127
5.5.C. Phosphorous isotopes
128
5.5.D. Sulfur isotopes
128
5.5.E. Iodine isotopes
129
5.6.
Kinetic isotope effects
129
5.7.
Isotope (hydrogen) exchange
130
5.7.
A. Mechanisms of exchange in model molecules
131
5.7.B. Monitoring exchange
135
1.
NMR
135
2.
Mass spectrometry
136
3.
Neutron diffraction
137
5.7.C. Exchange in macromolecules
137
1.
Solvent penetration model
137
2.
Local unfolding mechanism
138
a. EXl mechanism
138
b. EX2 mechanism
139
viii CONTENTS
6.
Mass spectrometry
140
6.1
Electrospray ionization (ESI)
142
6.2.
Matrix-assisted laser desorption/ionization (MALDI)
144
6.3.
Mass Analyzers
146
6.3.A. Magnetic focusing
146
6.3.B. Quadrupole mass analyzers
146
6.3.C. Time-of-flight analyzers)
147
6.3.D. Fourier-transform ion cyclotron resonance (FTMS)
148
6.4.
Tandem Mass Spectrometry (MSn)
148
Section II: Visualizing macromolecules
7.
Scattering of radiation by molecules
150
7.1.
Static light scattering
152
7. I.A.
Measuring the size of
a
macromolecule
152
7. LB.
Diffraction effects from the internal structure of a particle
154
7.
l.C. The radius of gyration
156
7.2.
X-ray scattering
158
7.2.
A. Small-angle scattering
161
1.
Fourier transforms and the vector length distribution
function, P(r)
162
2.
Determining the radius of gyration from small-angle
scattering data
164
7.2.
B. Interactions between particles
166
7.2.
C. Experimental apparatus
167
7.2.D. X-ray sources and detectors
168
7.3.
Neutron scattering
169
7.3.
A. Interactions of neutrons with atoms
169
7.3.B. Contrast variation
170
7.3.C. Contrast variation and small-angle scattering
171
7.3.D. Production and detection of neutrons
172
8.
Microscopy and scanning probes
174
8.1.
Basic principles of microscopy
175
8.2.
Light microscopy
177
8.2.
A. Transmission light microscopy
177
1.
Bright-field microscopy
177
2.
Dark-field microscopy
177
3.
Phase-contrast microscopy
178
4.
Polarization microscopy:
biréfringent
objects
179
5.
Differential interference-contrast (DIC) microscopy
179
8.2.B. Fluorescence microscopy
180
1.
Immunorluorescence microscopy
181
8.2.
С
Confocal microscopy
182
8.2.D. Near-field scanning optical microscopy (NSOM)
184
CONTENTS ix
8.3. Electron
microscopy
185
8.3.
Α.
Transmission
electron microscopy
(ТЕМ)
186
8.3.B. Negative stain
187
1.
Rotary shadowing (shadowcasting)
189
8.3.
С
Cryoelectron microscopy
189
8.3.D. Scanning electron microscopy
(SEM) 191
1.
Environmental scanning electron microscope
193
8.3.E. Scanning transmission electron microscopy (STEM)
193
8.3.F. Immunoelectron microscopy
195
8.3.G. Freeze-fracture microscopy
195
8.4.
Reconstructing
3-D
structures from 2-D microscopy
196
8.4.A. Single-particle reconstruction
196
8.4.B. Electron tomography
197
8.5.
Scanning probes
198
8.5.A. Scanning tunneling microscopy
199
8.5.B. Atomic force microscopy
201
1.
Principles
202
2.
Imaging methods
203
3.
The sample
204
4.
Forces detected
205
5.
Resolution
205
8.5.C. Magnetic force microscopy
206
8.6.
Manipulating individual molecules
207
9.
Crystallography
210
9.1.
Crystallization of macromolecules
211
9. I.A.
Precipitation of macromolecules
213
1. Sulfate
salts
213
2.
2-Methyl-2,4-pentanediol (MPD)
214
3.
PEG (polyethylene
glycol)
215
9.1.
В.
Vapor-phase crystallization
215
9.2.
X-ray crystallography
216
9.2.A. Crystallography in pictures: optical transforms
219
9.2.B. Bragg angle: reflecting X-rays
221
9.2.C. Reciprocal space
222
9.2.D. Precession photograph: revealing reciprocal space
223
9.2.E. Measuring the structure factors
223
9.2.F. Temperature factor: smearing the electron density
224
1.
Wilson plot
225
9.2.G. Cryocrystallography: preservation by freezing
226
9.3.
Determining the phases
227
9.3.
A Direct methods: phases from amplitudes
227
9.3.
В
Isomorphous replacement: heavy atoms
228
9.3.
С
Molecular replacement: an approximate solution
231
9.
3.D. Anomalous dispersion: scattering from the edge
232
9.3.E. Multiple-wavelength anomalous dispersion (MAD)
233
CONTENTS
9.4.
Calculating the electron density map
233
9.5.
Structure refinement
236
9.5.A. R-factor
238
9.5.B. Solvent flattening: using the background
239
9.5.C. Molecular averaging: noncrystallographic symmetry
240
9.6.
Difference Fourier: locating bound ligands or structural differences
240
9.7. Laue
diffraction
241
9.8.
Neutron diffraction
242
9.9.
Fiber diffraction: disorder in two dimensions
243
9.9.
Electron crystallography. 2-D crystals
245
Section III: Spectroscopy
10.
Absorption and emission of light
248
10.1.
Absorption spectroscopy: excited states
249
10.I.A.
Absorbance spectrophotometers
253
10.1.B. Absorbance properties of proteins
255
10.
l.C. Absorbance of nucleic acids
258
10.I.D.
Absorbance to determine concentrations
259
10.
I.E. Difference spectroscopy
260
10.1.F. Solvent perturbation spectroscopy: interactions of chromophores
with the solvent
260
10.1.
G. Linear dichroism
261
10.2.
Fluorescence spectroscopy: releasing the excitement
261
10.2.
A. Measuring fluorescence
264
10.2.B. Phosphorescence: glowing in the dark
265
10.2.
С
Fluorescence yields and life-times
266
10.2.
1). Fluoresence quenching: dampening the excitement
268
10.2.
E. Fluorescence polarization or anisotropy: exciting orientations
270
10.2.
F. Fluorescence of proteins
270
10.3.
Fluorescence energy transfer (FRET): sharing the excitement
272
10.4.
Reporter groups
275
10.5.
Luminescence
277
10.5.
A. Chemiluminescence
277
Ю.б.В.
Luciferins and luciferases
279
1.
Firefly luciferase
280
2.
Bacterial luciferases
281
3.
Aequorin and calcium-binding photoproteins
283
4.
Green fluorescent protein
284
10.6.
EXAFS (extended X-ray absorption fluorescence spectrophotometry)
287
11.
Circular dichroism
291
11.1.
The basics: chiral consequences
291
11.I.A.
Rotational strength
294
11.
l.B. Optical rotation: Cotton effects
294
11.2.
CD spectrophotometers
296
CONTENTS xi
11.3. CD
of proteins
298
11.3.
A. Secondary structure analysis
299
1.
α
-Helix
300
2. ß-Sheet 300
3. ß-Turns 301
4.
Irregular conformations
302
5.
Determining protein secondary structure
302
11.3.B. Tertiary structure of proteins
303
11.3.
С
Membrane proteins
304
11
.3.D. Binding of ligands
304
11.4.
CD of nucleic acids
305
11.4.
A. Secondary structure
307
1.
B-DNA
307
2.
A-DNA
308
3.
Z-DNA
308
4.
Correlation with helix sense
308
5.
Double-stranded
RNA
309
6.
Single-stranded
DNA
and
RNA
309
1
1.4.В.
Tertiary structure
309
1
1.4.С.
Binding of ligands to nucleic acids
309
11.5.
Fluorescence-detected CD
310
12.
Vibrational spectroscopy
311
12.1.
Molecular vibrations
312
12.2.
Vibrational spectra
314
12.2.
A. Infrared
(IR)
spectroscopy
315
12.2.
B. Raman spectroscopy
318
12.2.
С
Resonance Raman spectroscopy
321
12.2.
D. Chiroptical techniques
323
12.2.E. Surface-Enhanced Raman Spectroscopy
[SERS]
323
12.2.F.
Non-linear Optical Techniques
324
12.2.
G. Vibrational microscopy
324
12.3.
Vibrational spectra of proteins
325
12.3.
A. Polypeptide backbone vibrations; the amide bond
325
12.3.B.
Amino
acid side-chains
328
1.
Trp
residues
329
2.
Tyr
residues
331
3.
His residues
331
4.
Cys residues
331
12.3.
С
Ligands and chromophores bound to proteins
332
12.4.
Nucleic acids
333
12.5.
Lipids
337
12.6.
Kinetic studies
338
13.
Nuclear magnetic resonance (NMR)
339
13.1.
The basics
340
xii CONTENTS
13.1
.Α.
Relaxation
processes
343
13.1.
В.
Generating an NMR spectrum
345
13.
l.C. Chemical shift
347
13.I.D.
Scalar coupling, J-coupling, spin-spin coupling
352
13.
I.E. Magnetization transfer
356
13.1.F. Isotopes and multi-dimensional NMR spectra
356
1.
Isotope editing or filtering
358
2.
NMR to monitor isotope exchange
359
13.2.
Solid-state NMR
359
13.3.
Resonance assignments to atoms
362
13.3.A. COSY spectrum
363
13.3.B. TOCSY spectrum
364
13.3.C. Nuclear Overhauser Effect
(NOE)
365
1.
NOESY spectrum
366
2.
ROESY spectrum
367
13.3.D. Sequential assignments
368
13.3.E. Residual dipolar couplings
369
13.4.
Generating structures from NMR data
371
13.4.
A. Distance geometry
373
13.4.B. Simulated annealing
374
13.5.
Structure simulations
375
13.5.
A. Potential functions (force fields)
376
13.5.B. Molecular mechanics and energy minimization
378
13.5.
C. Molecular dynamics
379
1.
Brownian dynamics
380
2.
Normal mode analysis
381
13.5.
D. Monte Carlo calculations
381
13.5.
E. Free energy calculations
382
1.
Statistical mechanical averaging
383
2.
Thermodynamic cycles
383
3.
Free energy perturbation
384
14.
Electron magnetic/paramagnetic/spin resonance
386
14.1.
The fundamentals
387
14.I.A.
Measuring an electron paramagnetic resonance (EPR) spectrum
388
14.
l.B. Theg-value
389
14.
l.C. Hyperfme structure
389
14.1.D. Anisotropy
390
14.
I.E. Spin-spin interactions
391
14.1.
F. Higher-order techniques
392
1.
Electron nuclear double resonance (ENDOR)
392
2.
Electron spin echo
(ESE)
393
3.
ESE
envelope modulation (ESEEM)
393
4.
ESE-ENDOR
393
CONTENTS xiii
14.2.
Stable
free radicals
393
14.2.
A. Nitroxides
394
14.2.
B. Nitric oxide complexes
395
14.2.
С
Paramagnetic metal ions
395
14.3.
Transient intermediates with unpaired electrons
397
14.3.A. Photosynthesis intermediates
397
14.3.B. Free-radical protein intermediates: ribonucleotide reductases
399
14.4.
Spin trapping and EPR imaging
400
14.5.
Spin labeling
402
14.5.A. Site-directed spin labeling (SDSL)
402
Section IV: Transport in solution
15.
Hydrodynamics: movements of molecules in solution
404
15.1.
Volumes of macromolecules in solution
404
15.I.A.
Hydrodynamic volume
407
15.2.
Excluded volume
409
15.3.
Translational diffusion
411
15.3.A. Dynamic light scattering (photon correlation spectroscopy,
quasi-elastic light scattering)
416
1.
Fluorescence correlation spectroscopy
419
15.3.B. How the structure affects the translational diffusion coefficient
420
1.
Hydrodynamic radius (Stokes radius)
420
2.
Frictional coefficient
421
3.
Frictional ratio
421
4.
Shape
422
5.
Molecular weight
424
15.3.C. Diffusion properties of macromolecules
425
15.4.
Rotational diffusion
426
15.4.
A. Fluorescence anisotropy decay
426
1.
Steady-state measurements
426
2.
Kinetic measurements
428
15.4.B. Electric birefringence decay
429
15.4.C. NMR relaxation
430
15.4.D. Observed rates of rotation
430
15.5.
Diffusion of small molecules through biomolecular systems
433
15.6.
Proteins diffusing in membranes
433
15.7.
Viscosity
434
16.
Sedimentation by centrifugation
438
16.1.
Preparative centrifugation
440
16.I.A.
Selective pelleting on the basis of size: differential centrifugation
441
16.2.
Analytical ultracentrifugation
441
16.2.
A. Instrumentation
443
1.
Optical systems
444
xiv CONTENTS
16.3. Sedimentation
velocity centrifugation
446
16.3.
Α.
Sedimentation
coefficient
(s -value)
446
1.
Fitting the Lamm equation to individual scans
448
2.
Analysis of the distribution of sedimentation coefficients
449
3.
Nonideality and
intermolecular
interactions
449
4.
Zone sedimentation
451
16.3.B. Measuring the diffusion coefficient directly
451
16.3.
С
Sedimentation velocity data to determine the molecular weight
and shape
452
1.
Molecular weight averages: monodispersity, polydispersity,
and paucidispersity
454
16.4.
Sedimentation equilibrium centrifugation
455
16.4.
A. Interactions between molecules
458
16.5.
Density-gradient centrifugation
458
16.5.
A. Materials used for density gradients
460
16.5.
B. Analyzing density gradients after centrifugation
461
1.
Rotors for density gradient separations
461
16.5.
С
Sedimentation velocity gradients
462
1.
Sucrose gradients
462
2.
Flotation
463
16.5.D. Isopycnic gradient centrifugation
463
17.
Electrophoresis
465
17.1.
Gel electrophoresis
467
17.I.A.
Polyacrylamide: free-radical polymerization
469
17.1.
B.
Agarose:
gelation and large pores
472
17.1
.C. Varying the gel concentration: Ferguson plot
473
17.I.D.
Fore gradient electrophoresis: dead ends
474
17.
I.E. Pulsed-field gel electrophoresis (PFGE): changing directions
475
17.1.
E Electroendosmosis
475
17.2.
Disc electrophoresis: buffer discontinuities
476
17.2.
A. Keeping track: tracking dyes
479
17.2.B. Isotachophoresis: stacking
479
17.3.
Affinity electrophoresis
481
17.3.
A. Immunoelectrophoresis
482
17.4.
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)
483
17.5. Hydrophobie
electrophoresis
485
17.6.
Isoelectric focusing (IEF)
486
17.6.
A. Carrier ampholytes: soluble amphoteric buffers
487
17.6.B. Immobilized
pH
gradients (IPG)
488
17.7.
Two-dimensional (2-D) gel electrophoresis
489
17.7.
A. Blue native PAGE
491
17.8.
Transverse gradient gel electrophoresis (TGGE)
491
17.8.
A. Isomerization during electrophoresis
493
17.8.B. Urea-gradient gel electrophoresis
495
17.8.
С
Temperature-gradient gel electrophoresis
496
CONTENTS xv
17.9.
Capillary electrophoresis
498
17.9.
A. Particle electrophoresis
499
17.10.
Sample detection and quantification
500
17.1 O.A.
Silver stain
501
17.10.B. Zymography: visualizing enzyme activities
503
17.10.C. Electroelution
505
18.
Molecular sieves: gel filtration/size exclusion chromatography
506
18.1.
Molecular sieve materials
507
18.1.A. Sephadex
510
18.1.B. Sepharose
511
18.1.C. Sephacryl
511
18.2.
Molecular sieving
511
18.3.
Size exclusion chromatography (SEC)
513
18.3.A. Small-zone filtration
514
18.3.
B. Large zones: boundary analysis
515
18.4.
Estimating the sizes of molecules
516
18.4.
A. Estimating the molecular weight
517
18.5.
Interacting molecules
518
18.6.
Isomerizing molecules
520
Section V. Interactions between molecules
19.
Ligand binding
522
19.1.
Determining the structures of bound ligands
523
19.I.A.
Difference electron density maps
523
19.1
.B. Nuclear magnetic resonance (NMR)
524
19.2.
Energetics of binding interactions
525
19.2.
A. Binding affinities: measuring the attraction
526
19.2.B. Group contributions to binding: dissecting the ligand
528
19.2.C. Thermodynamics of association
533
19.2.D. Rates of binding
534
19.2.E. One-and two-dimensional binding
537
19.3.
Methods to measure binding
537
19.3.A. Filtration: separating ligand and
macromolecule
538
19.3.B. Ultracentrifugation
538
19.3.C. Equilibrium dialysis:
semipermeable
membranes
538
1.
Donnan
effect
540
2.
Flow dialysis
540
3.
Dialysis: changing the solvent
541
4.
Osmotic pressure due to the
macromolecule
541
19.3.
D.
Titration
microcalorimetry: the heat ot binding
542
19.3.E. Displacement of a labeled ligand
543
19.3.F. Fluorescence polarization and correlation spectroscopy
543
19.4.
Graphical analysis of binding data
543
19.4.A. Direct plot
544
19.4.B. Semilogarithmic plot
545
xvi CONTENTS
19.4.C. Double-reciprocal
plot
545
19.4.D. Scatchard
plot
545
19.4.E. Hill
plot
547
19.5. Multiple
binding sites: the possibility of cooperativity
547
19.5.
A. Identical and independent sites
547
19.5.B. Heterogeneous sites or interactions between identical sites
548
19.5.C. Direct plots
548
19.5.D. Semi-logarithmic plots
548
19.5.E. Double-reciprocal plots
548
19.5.E Non-linear Scatchard plots
549
19.5.G. Non-linear Hill plots
551
19.5.
H. Interactions between
différent ligands:
linked functions
551
19.6.
Affinity labeling
553
19.7.
Light-activated (caged) ligands
555
19.8
Cross-linking
557
19.8.A. Bifunctional cross-linking reagents
557
1.
Glutaraldehyde
560
19.8.B. Cross-linking procedures
560
19.8.
С
Applications of cross-linking
561
20.
Chromatography
562
20.1.
Paper and thin layer chromatography: planar chromatography
564
20.2.
Column-liquid chromatography
565
20.2.
A. Elution methods
566
1.
Gradient elution
567
2.
Isocratic elution
567
20.2.
B. Ion-exchange chromatography
567
1.
Chromatofocusing
570
20.2.
C. Reversed-phase chromatography
572
20.2.
D. Hydrophilic interaction chromatography
572
20.2.
E.
Hydrophobie
chromatography
573
20.2.
E Hydroxyapatite
chromatography
576
20.2.
G. Affinity chromatography
577
1.
Coupling of the ligand to an inert carrier
578
2.
Selective adsorption and elution of the desired
macromolecule
581
3.
Limitations to biospecificity
582
20.2.
H. Immunoaffinity chromatography
584
20.3.
Gas-liquid chromatography
584
21.
Interactions of immobilized macromolecules
586
21.1.
Blotting: immobilizing macromolecules
586
21.2.
Blotting matrices: sticky filters
587
21.2.
A. Nitrocellulose
588
21.2.
B. Nylon membranes
589
21.2.C. Poly(vinylidene fluoride) (PVDF)
590
CONTENTS xvii
21.3.
Blotting
procedures
590
21.3.A.
Dot blots
590
21.
3.В.
Gelblots 591
1.
Diffusion blotting
591
2.
Convection blotting
591
3.
Electroblotting
591
21.3.C. Colony blots
592
21.4.
Blot probes: locating the missing
macromolecule
592
21.4.
Α.
DNA
probes
593
1.
Southern blots
(DNA
blots)
593
2.
Northern blots
(RNA
blots)
594
21.4.B. Western blots (protein blots)
595
1.
Lectin blotting for carbohydrates
596
2.
Ligand blotting
597
3.
Cell blotting
597
21.5.
Reporting systems
597
21.5.A. Chromogenic enzyme substrates
597
1.
Alkaline phosphatase
598
2.
Peroxidases
599
21.5.B. Avidin-biotin system
600
21.6
ELISA
(Enzyme-linked immunosorbent assay)
604
21.6.
A. Competitive
ELISA
605
21.6.B. Indirect
ELISA
606
21.6.
С.
Sandwich assay
606
21.7.
Microarrays
608
21.8.
Surface plasmon resonance: changes in reflectance
609
|
| any_adam_object | 1 |
| author | Creighton, Thomas E. |
| author_facet | Creighton, Thomas E. |
| author_role | aut |
| author_sort | Creighton, Thomas E. |
| author_variant | t e c te tec |
| building | Verbundindex |
| bvnumber | BV036613823 |
| classification_rvk | WD 2200 WD 4150 |
| classification_tum | CHE 802f PHY 821f BIO 220f |
| ctrlnum | (OCoLC)682039359 (DE-599)BVBBV036613823 |
| dewey-full | 572.83 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 572 - Biochemistry |
| dewey-raw | 572.83 |
| dewey-search | 572.83 |
| dewey-sort | 3572.83 |
| dewey-tens | 570 - Biology |
| discipline | Physik Biologie Chemie |
| format | Book |
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| id | DE-604.BV036613823 |
| illustrated | Illustrated |
| indexdate | 2024-07-09T22:44:12Z |
| institution | BVB |
| isbn | 9780956478108 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-020534011 |
| oclc_num | 682039359 |
| open_access_boolean | |
| owner | DE-M49 DE-BY-TUM DE-355 DE-BY-UBR DE-19 DE-BY-UBM DE-91G DE-BY-TUM |
| owner_facet | DE-M49 DE-BY-TUM DE-355 DE-BY-UBR DE-19 DE-BY-UBM DE-91G DE-BY-TUM |
| physical | XXXIX, 642 S. Ill., graph. Darst. |
| publishDate | 2010 |
| publishDateSearch | 2010 |
| publishDateSort | 2010 |
| publisher | Helvetian Press |
| record_format | marc |
| spelling | Creighton, Thomas E. Verfasser aut The physical and chemical basis of molecular biology Thomas E. Creighton Molecular biology [Eastbourne] Helvetian Press 2010 XXXIX, 642 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Molekulare Biophysik (DE-588)4170391-1 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 gnd rswk-swf Biochemie (DE-588)4006777-4 gnd rswk-swf Molekularbiologie (DE-588)4039983-7 s Molekulare Biophysik (DE-588)4170391-1 s Biochemie (DE-588)4006777-4 s b DE-604 Digitalisierung UB Regensburg application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020534011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Creighton, Thomas E. The physical and chemical basis of molecular biology Molekulare Biophysik (DE-588)4170391-1 gnd Molekularbiologie (DE-588)4039983-7 gnd Biochemie (DE-588)4006777-4 gnd |
| subject_GND | (DE-588)4170391-1 (DE-588)4039983-7 (DE-588)4006777-4 |
| title | The physical and chemical basis of molecular biology |
| title_alt | Molecular biology |
| title_auth | The physical and chemical basis of molecular biology |
| title_exact_search | The physical and chemical basis of molecular biology |
| title_full | The physical and chemical basis of molecular biology Thomas E. Creighton |
| title_fullStr | The physical and chemical basis of molecular biology Thomas E. Creighton |
| title_full_unstemmed | The physical and chemical basis of molecular biology Thomas E. Creighton |
| title_short | The physical and chemical basis of molecular biology |
| title_sort | the physical and chemical basis of molecular biology |
| topic | Molekulare Biophysik (DE-588)4170391-1 gnd Molekularbiologie (DE-588)4039983-7 gnd Biochemie (DE-588)4006777-4 gnd |
| topic_facet | Molekulare Biophysik Molekularbiologie Biochemie |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=020534011&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT creightonthomase thephysicalandchemicalbasisofmolecularbiology AT creightonthomase molecularbiology |