Practical Protein Chemistry - A. Darbre 1989
Determination of the composition of protein oligomers. Preparation of monomers and polypeptide chains
Methods for identification of oligomers
Determination of protein oligomer composition. Preparation of monomers and polypeptide chains
Methods for oligomer identification
Determination of the composition of protein oligomers. Preparation of monomers and polypeptide chains
Methods for determining the molecular weight of monomers
Methods for isolation of monomers
Stoichiometric ratio of monomers in the oligomer
Determination of oligomer composition from molecular weights of monomers
Cross-linking of subunits with bifunctional reagents
Polyacrylamide gel electrophoresis
Determination of the composition of protein oligomers. Isolation of monomers and polypeptide chains
Cross-linking of oligomer and monomer
Cross-linking of oligomers and monomers
Other types of cross-links in proteins
Determination of the composition of protein oligomers. Preparation of monomers and polypeptide chains
Preparation of monomers for sequencing
Hydrolysis of N-terminal pyroglutamic acid
Reduction of sulfoxides with N-methylmercaptoacetamide
Techniques for working with lipoproteins
Cleavage of oligosaccharide fragments from glycoproteins
Determination of protein oligomer composition. Preparation of monomers and polypeptide chains
Chemical fragmentation of polypeptides
Cleavage of disulfide bonds
Reduction and S-carboxymethylation
Chemical Fragmentation of Polypeptides
Cleavage of Disulfide Bonds
Other Protecting Groups (for the SH group)
Polypeptide Fragmentation by Chemical Methods
Cleavage of S-S Groups by Oxidation
Chemical fragmentation of polypeptides
Cleavage of disulfide bonds
Chemical Fragmentation of Polypeptides
Polypeptide Fragmentation by Chemical Methods
Chemical Fragmentation of Polypeptides
Polypeptide fragmentation by chemical methods
Cleavage at the methionine residue
Polypeptide Fragmentation by Chemical Methods
Cleavage at Methionine Residues
Cleavage at Tryptophan Residues
Chemical Fragmentation of Polypeptides
Cleavage at Tryptophan Residues in the DMSO – Hydrohalic Acid System
Chemical fragmentation of polypeptides
Cleavage at tryptophan residues
Cleavage at tryptophan residues using cyanogen bromide in heptafluorobutyric acid
Chemical Fragmentation of Polypeptides
Cleavage at Tryptophan Residues
Cleavage at Tryptophan Residues with o-Iodosobenzoic Acid
Polypeptide Fragmentation by Chemical Methods
Polypeptide fragmentation by chemical methods
Cleavage at the tryptophan residue
Polypeptide Fragmentation by Chemical Methods
Cleavage at Tyrosine Residues
Chemical fragmentation of polypeptides
Cleavage at tyrosine residues
Chemical Fragmentation of Polypeptides
Cleavage at Tyrosine Residues
Polypeptide Fragmentation by Chemical Methods
Cyanylation with 2-Nitro-5-thiocyanobenzoic Acid
Chemical Fragmentation of Polypeptides
Other Methods of Chemical Peptide Bond Cleavage
Other Methods of Chemical Cleavage of Peptide Bonds
Polypeptide Fragmentation by Chemical Methods
Other Methods of Chemical Peptide Bond Cleavage
Cleavage at the Dehydroalanine Residue
Other Methods of Chemical Cleavage of Peptide Bonds
Other Methods of Peptide Bond Cleavage
Chemical Fragmentation of Polypeptides
Chemical fragmentation of polypeptides
Enzymatic fragmentation of the polypeptide chain
General conditions for enzymatic hydrolysis
Duration of enzymatic hydrolysis
Monitoring the enzymatic hydrolysis process
Termination of the enzymatic reaction
Protein modification methods
Alkylation of sulfhydryl groups
Fragmentation of the polypeptide chain by enzymatic methods
Modification of lysine residues
Enzymatic fragmentation of the polypeptide chain
Modification of arginine residues
Methods for protein modification
Modification of protein carboxyl groups
Fragmentation of the polypeptide chain by enzymatic methods
Proteases with high specificity
Enzymatic fragmentation of the polypeptide chain
High-specificity proteases
Fragmentation of the polypeptide chain by enzymatic methods
Proteases with high specificity
V8 Protease from Staphylococcus aureus
Fragmentation of the Polypeptide Chain by Enzymatic Methods
Proteases with High Specificity
Enzymatic fragmentation of the polypeptide chain
Proteases with high specificity
Protease from Armillaria mellea
Protease II from Myxobacter AL1
Fragmentation of the polypeptide chain by enzymatic methods
Enzymatic fragmentation of the polypeptide chain
Proteases with low specificity
Fragmentation of Polypeptide Chains by Enzymatic Methods
Proteases with Low Specificity
Enzymatic fragmentation of the polypeptide chain
Proteases with low specificity
α-Lytic protease from Sorangium sp.
Disulfide Bonds
Schematic Diagram of the Analysis
Determination of the Number of Disulfide Bonds
Disulphide bonds
Cleavage of the polypeptide chain into short cystine-containing peptides
Disulfide Bonds
Hydrolysis by trypsin and chymotrypsin
Hydrolysis by Staphylococcal Protease
Cleavage of the Polypeptide Chain into Short Cystine-Containing Peptides
Cleavage of the polypeptide chain into short cystine-containing peptides
Hydrolysis following maleylation and succinylation
Fractionation of Cystine-Containing Peptides
Detection of Cystine-Containing Peptides
Identification and Localization of Cystine-Containing Peptides
Disulfide bonds
Identification and localization of cystine-containing peptides
Identification based on a known amino acid sequence
Disulfide Bonds
Identification of disulfide bonds in proteins with unknown amino acid sequences
Disulfide bonds
Disulfide Bonds
Special Methods
Disulfide bonds
Special methods
Disulfide Bonds
Special Methods
Specific Cleavage at Cysteine Residues
Affinity Chromatography of Proteins
Applications of Affinity Chromatography
Methods for Preparation of Affinity Sorbent
Methods for Preparing Affinity Sorbents
General Procedures in Affinity Chromatography
General Techniques of Affinity Chromatography
General Procedures in Affinity Chromatography
Immobilized Substrates, Inhibitors, and Enzyme Cofactors
Examples
Isolation and Purification of Antibodies
Purification of Hormone Receptors
Purification Strategy for Rare Proteins
Separation of Protein and Peptide Mixtures by High-Performance Liquid Chromatography
Separation of protein and peptide mixtures by high-performance liquid chromatography
Gel filtration
Gel chromatography
Determination of true molecular weights
Adsorption and partition of polypeptides and proteins
Sample application preparation
Selection of columns for reversed-phase separation
Reversed-phase separation of peptides
Selection of mobile phase composition
Isolation of peptides using phosphoric acid
Peptide separation using trifluoroacetic and heptafluorobutyric acids
Reversed-phase peptide separation
Correlation between retention and structure
High-speed liquid chromatography of proteins
Peptide Mapping of Proteins
Peptide mapping of proteins
Peptide mapping — types of structures analyzed
Peptide mapping in practice
Two-dimensional peptide mapping
Peptide Mapping of Proteins
Peptide Mapping in Practice
Peptide Mapping by High-Performance Liquid Chromatography
Peptide mapping of proteins
Analytical Methods
Concentration of Protein Solutions
Exhaustive protein hydrolysis for amino acid analysis
Tryptophan
Determination of tryptophan following alkaline protein hydrolysis
Determination of tryptophan following acid hydrolysis of protein
Determination of Tryptophan in Intact Proteins
Dicarboxylic amino acid amide residues
Methods for Amide Determination
Sulfur-containing amino acids
Arginine
Phosphorylated Amino Acids
y-Carboxyglutamic Acid
y-Carboxyglutamic acid
Analytical methods
Analytical Methods
Determination of acetyl and formyl groups
Detection of compounds in paper chromatography and thin-layer chromatography
Detection of compounds in paper and thin-layer chromatography
Thin-Layer Chromatography of Amino Acids
Amino Acid Column Chromatography
Gas-Liquid Chromatography of Amino Acids
General Sample Preparation Procedure
Quantitative Protein Determination
Protein Determination Using the Folin–Ciocalteu Reagent
Protein Determination Using the Biuret Reaction
Protein Determination Using Coomassie Brilliant Blue
Protein Determination Using Sulfobromophthalein
Protein Determination Using Trinitrobenzenesulfonic Acid in the Presence of Lipids
Atomic Absorption Spectrophotometry Method
Methods for Staining Proteins in Gels
Protein Staining Methods Using Dyes
Protein Staining Methods in Gels
Protein Staining with Silver Complexes
Protein Staining on Nitrocellulose Paper
Detection Using Silver and Antibody Complexes
Determination of Protein Bound to a Solid Support
Determination of protein bound to a solid support
Determination of amino groups on a solid support
Determination of protein attached to a solid support
Determination of amino groups on hydrophilic matrices
Determination of amino groups by reaction with chloranil
Determination of Protein Bound to a Solid Support
Determination of the Nature of the Resin Substituent by Staining
Determination of protein attached to a solid support
Colorimetric determination of carbodiimides
Fluorometric determination of carbodiimide
Glycoproteins
Carbohydrate-Containing Proteins
Methyl Glycoside Trimethylsilyl Method
Sensitive Methyl Glycoside Trimethylsilyl Method
Gas Chromatography-Mass Spectrometry Method
Determination of neutral sugars and aminosugars as alditol acetates by GLC
Glycoproteins
Determination of alditols by thin-layer and gas-liquid chromatography
Carbohydrate-Containing Proteins
Determination of neutral sugars and aminosugars on an amino acid analyzer
Enantiomeric analysis of amino acid mixtures by high-performance liquid chromatography
Reagents
Synthesis of N, N-di-n-propyl-L-alanine
Traditional strategy for protein structure determination
Fractionation of soluble peptides
Traditional strategy for determining protein structure
Traditional Strategy for Protein Structure Determination
Fractionation of Soluble Peptides
Dowex-50 Column Chromatography
Traditional strategy for determining protein structure
Fractionation of soluble peptides
DEAE-cellulose column chromatography
High-voltage electrophoresis and paper chromatography
Traditional strategy for protein structure determination
Fractionation of insoluble peptides
Homogeneity control and determination of the amino acid sequence of peptides
Reconstruction of the polypeptide chain
Methods
Preparative electrophoresis and paper chromatography
Traditional strategy for determining protein structure
Traditional strategy for protein structure determination
Determination of the amino acid sequence of peptides by the Edman degradation method with identification of dansyl amino acids
Determination of the amino acid sequence of peptides by the Edman method with identification of dansyl amino acids
Edman condensation and cleavage
Edman degradation. Methodology
Methods for Solid-Phase Amino Acid Sequence Analysis
Materials
Methods of solid-phase amino acid sequence analysis
Methods for Solid-Phase Amino Acid Sequence Analysis
Glass beads for diluting polymer samples
Reagent and Buffer Solutions for the Sequenator
Methods of Solid-Phase Amino Acid Sequence Analysis
Resin Synthesis
Methods for Solid-Phase Amino Acid Sequence Analysis
Methods of Solid-Phase Amino Acid Sequence Analysis
Determination of Amino Group Content in Supports Based on Organic Polymers or Porous Glasses
Methods for Solid-Phase Amino Acid Sequence Analysis
Coupling Methods
Coupling of Lysyl-Containing Peptides Using DITC
Methods of solid-phase amino acid sequence analysis
Attachment methods
DITC method using isothiocyanate glass
Methods of solid-phase amino acid sequencing
Coupling methods
Coupling via homoserine lactone
Methods of solid-phase amino acid sequence analysis
Carboxyl coupling using carbodiimide
Monitoring coupling efficiency
Methods of solid-phase amino acid sequencing
Cleavage on a solid-phase sequencer
Methods of solid-phase amino acid sequence analysis
Methods for Solid-Phase Amino Acid Sequence Analysis
Cleavage on a Solid-Phase Sequenator
Sequenator Operational Program
Cleavage in a Solid-Phase Sequenator
Cleavage on a Solid-Phase Sequenator
Cleavage of Anilinothiazolinone
Rearrangement of Anilinothiazolinone into PTH
Discussion
Solid-Phase Analysis of Small Peptides
Solid-Phase Analysis of Large Peptides and Proteins
Comparison of Liquid-Phase and Solid-Phase Methods
Analysis of Amino Acid Phenylthiohydantoins
Analytical Methods
High-Performance Liquid Chromatography
Analysis of amino acid phenylthiohydantoins
Analysis of Amino Acid Phenylthiohydantoins
Analysis of amino acid phenylthiohydantoins
Back-hydrolysis of PTH-amino acid derivatives
Analysis of Amino Acid Phenylthiohydantoins
Determination of amino acid sequence using 4-dimethylaminoazobenzene-4'-isothiocyanate (manual procedure)
Determination of Amino Acid Sequence Using 4-Dimethylaminoazobenzene-4'-isothiocyanate (Manual Procedure)
Determination of amino acid sequence using 4-dimethylaminoazobenzene-4'-isocyanate (manual procedure)
DABITC-FITC liquid-phase method
DABITC–FITC solid-phase methodology
Determination of amino acid sequence using 4-dimethylaminoazobenzene-4'-isothiocyanate (manual procedure)
Conversion of amino acid thiazolinones to thiohydantoins
Identification of DABTC-amino acids
Amino acid sequence determination using 4-dimethylaminoazobenzene-4'-isothiocyanate (manual procedure)
Identification of DABTC amino acids
High-Performance Liquid Chromatography
Determination of Amino Acid Sequence Using 4-Dimethylaminoazobenzene-4'-isothiocyanate (Manual Procedure)
Current state of automated liquid-phase amino acid sequence analysis
Comparison of Quadrol with volatile buffers
Key challenges encountered when using an automated sequencer
Major problems encountered when using an automated sequencer
Vacuum line leaks in a single-pump system
Major challenges encountered when using an automated sequencer
Main problems arising from the use of an automated sequencer
Major problems encountered when using an automated sequencer
Instrument performance quality control
The latest methods for solid-phase and liquid-phase amino acid sequence determination
Latest methods of solid-phase and liquid-phase amino acid sequence determination
Solid-Phase Analysis. Novel Approaches
The latest methods of solid-phase and liquid-phase amino acid sequence determination
Solid-Phase Analysis. The Latest Approaches
Latest methods for solid-phase and liquid-phase amino acid sequence determination
Solid-Phase Analysis: The Latest Approaches
Attachment of peptides to supports and "blocking" of the remaining reactive sites of the support
The latest methods for solid-phase and liquid-phase determination of amino acid sequences
Latest methods for solid-phase and liquid-phase amino acid sequence determination
Solid-Phase Analysis. Recent Approaches
Other Sequential Degradation Methods
The latest methods for solid-phase and liquid-phase amino acid sequence determination
Automatic liquid-phase analysis. Improved procedures
Analysis of proteins and large peptides at the conventional level
Latest methods for solid-phase and liquid-phase amino acid sequence determination
Automated liquid-phase analysis. Advanced procedures
Amino acid sequence analysis of small and hydrophobic peptides
Latest methods of solid-phase and liquid-phase amino acid sequence determination
Automatic liquid-phase analysis. Advanced techniques
Latest methods for solid-phase and liquid-phase amino acid sequence determination
State-of-the-art methods for solid-phase and liquid-phase amino acid sequencing
Amino acid sequence analysis at the micro level using a gas-phase peptide-protein sequencer
Micro-scale amino acid sequence analysis using a gas-phase peptide-protein sequencer
Microscale amino acid sequence analysis using a gas-phase peptide-protein sequencer
Micro-scale amino acid sequence analysis using a gas-phase peptide-protein sequencer
Polypeptide purification for microanalysis
Instrumentation and Methodology
Discussion
Sample immobilization and instrument sensitivity
Microscale amino acid sequence analysis using a gas-phase peptide-protein sequencer
Micro-scale amino acid sequence analysis using a gas-phase peptide-protein sequencer
Determination of C-terminal amino acid sequence
Isolation and identification of C-terminal peptides
Methods of ion-exchange chromatography
Determination of C-Terminal Amino Acid Sequence
Isolation and Identification of C-Terminal Peptides
Two-Dimensional Peptide Mapping
Determination of C-Terminal Groups
Determination of the C-terminal amino acid sequence
Determination of C-terminal groups
Determination of C-terminal amino acid sequence
Determination of C-terminal amino acids as aldehydes
Determination of C-terminal amino acids via oxazolone alcoholysis
Determination of the C-terminal amino acid sequence
Determination of the C-terminal sequence
Cleavage by means of thiocyanate
Determination of C-terminal amino acid sequence
Determination of C-terminal sequence
Other chemical cleavage methods
Determination of the C-terminal amino acid sequence
Determination of the C-terminal sequence
Determination of C-Terminal Amino Acid Sequence
Determination of the C-terminal amino acid sequence
Application of electron impact mass spectrometry for determining the amino acid sequence of peptides and proteins
Requirements for the analyzed peptides
Requirements for analytical instruments
Interpretation of Mass Spectra
Interpretation of mass spectra
Peptide fragmentation pathways
Interpretation of Mass Spectra
Special cases of application of the method
Peptides with a blocked N-terminal amino group
Application of Electron Impact Mass Spectrometry for Determining the Amino Acid Sequence of Peptides and Proteins
Special Cases of Method Application
Determination of the N-Terminal Amino Acid Sequence
Application of electron impact mass spectrometry for determining the amino acid sequence of peptides and proteins
Special cases of method application
Proteins containing γ-carboxyglutamic acid residues
Application of Electron Impact Mass Spectrometry for Determining the Amino Acid Sequence of Peptides and Proteins
Application of Electron Impact Mass Spectrometry to Determine the Amino Acid Sequence of Peptides and Proteins
X-Ray Crystallography and Electron Microscopy
X-Ray Diffraction
X-ray crystallography and electron microscopy
Molecular imaging as a method of structural analysis
X-ray diffraction
Imaging of molecules: a type of structural analysis
Principles of structure determination in X-ray structural analysis
X-ray Crystallography and Electron Microscopy
X-ray Diffraction
X-Ray Crystallography and Electron Microscopy
X-Ray Diffraction
X-ray Crystallography and Electron Microscopy
X-ray Diffraction
Computer-Aided Molecular Modeling
Electron Microscopy
X-ray crystallography and electron microscopy
Electron microscopy
Resolution and limitations of the method
X-ray Crystallography and Electron Microscopy
Electron Microscopy
Sample Preparation and Staining
X-ray crystallography and electron microscopy
Electron microscopy
X-ray Crystallography and Electron Microscopy
Electron Microscopy
Three-Dimensional Electron Microscopy
X-ray crystallography and electron microscopy
Prediction of Peptide and Protein Conformation
Limitations of Traditional Methods
An Arsenal of Modern Theoretical Methods
The Arsenal of Modern Theoretical Methods
An Arsenal of Modern Theoretical Methods
The Arsenal of Modern Theoretical Methods
Periodic Evaluation and Mapping
An Arsenal of Modern Theoretical Methods
Secondary Structure Prediction. Special Calculation Methods
Basic Assumptions for Calculations
Basic premises of calculations
Testing of potential functions derived from crystallographic data using simple approximate methods
Initial Prerequisites for Calculations
Testing of Potential Functions Derived from Structural Data Using More Precise Methods
Basic Assumptions of Calculations
Attempts to Predict the Tertiary Structure of Globular Proteins