The development of innovative, efficient ionization techniques allowing a gentle desorption of even complex molecules has provided mass spectrometry with the breakthrough in the direct analysis of biopolymers. Applicable in various disciplines and yet more powerful than almost any other method, the spectrum of compounds investigated ranges from macromolecular chemistry via biochemistry right up to molecular biology. In this way, suitable mass spectroscopic techniques determine not only the primary structure of proteins but also provide an insight into their spatial structure, even allowing structural and functional analyses. The simplicity of electrospray MS for analyzing non–covalent biomacromolecular complexes opens up brand new perspectives for supramolecular chemistry as well as molecular recognition processes. The combination of efficient separation techniques such as affinity chromatography or new methods directly monitoring biomolecular interactions (biosensors) with mass spectrometry provides the basis for diagnotic platforms in clinical applications.
In his book, the internationally renowned expert Michael Przybylski provides a comprehensive overview of this still relatively young methodology, which will soon be as much an integral part of biopolymer structure analysis as X–ray crystallography and NMR spectroscopy.
In his book, the internationally renowned expert Michael Przybylski provides a comprehensive overview of this still relatively young methodology, which will soon be as much an integral part of biopolymer structure analysis as X–ray crystallography and NMR spectroscopy.
Table of Contents
INTRODUCTION: FUNDAMENTALS OF BIOPOLYMER MASS SPECTROMETRYHistory and instrumental development of biopolymer mass spectrometry
Analytical preconditions for mass spectrometry of macromolecules
Scope of interdisciplinary application areas
IONIZATION METHODS FOR BIOPOLYMER MASS SPECTROMETRY
Desorption– Ionisation Methods for Biopolymer Ion Formation –
Initial developments
Field Desorption
252–Cf Plasma Desorption & Fast Atom Bombardment
Matrix–Assisted Laser Desorption– Ionisation (MALDI)
Electrospray– Ionisation (ESI)
New Developments of Ionisation Techniques
INSTRUMENTATION FOR BIOPOLYMER MASS SPECTROMETRY
Analysers for Macromolecular Mass Spectrometry
Magnetic Sector Analysers
Quadrupole Mass Analysers
Time–of–Flight Mass Analysers
Ion Trap Mass Analysers
Hybrid Analysers
High Resolution Mass Spectrometers for Biopolymers
Fouriertransform– ICR Mass Spectrometry
Orbitrap Mass Spectrometry
Ion Detection and Spectra Acquisition
Preconditions for Macromolecular Ion Detectors
Spectra acquisition & data evaluation methods
Special High Mass Ion Detectors
SEPARATION TECHNIQUES AND HYBRID SYSTEMS FOR BIOPOLYMER MASS SPECTROMETRY
Liquid Chromatography– Mass Spectrometry
Size Exclusion Chromatography
Combination with Gel and Capillary Electrophoresis
Bioaffinity– Mass Spectrometry
Ion Mobility Mass Spectrometry
Online Biosensor– MS
METHODS FOR STRUCTURE DETERMINATION OF BIOPOLYMERS
Molecular Weight Determination
Mechanisms of Biopolymer Ion Formation
Mass Resolution and Accuracy of Macromolecular Ions
Methods for Primary Structure Determination of Polypeptides
Fragmentation of Polypeptide Ions for Tandem–MS Sequence Determination
Chemical and Enzymatic Degradation for Sequence Determination
Approaches for "Top–Down" Sequence Determination
Structural Characterization of Polypeptides by Chemical Modification Combined with Mass Spectrometry
Chemical Modification Reactions for Structure Analysis
Structural Characterisation using Crosslinking Reactions
Analysis of Non–covalent Supramolecular Biopolymer Complexes
Preconditions for Analysis of Non–covalent Interactions
Specificity Analysis of Biopolymer Complexes
Correlation of Solution and Gas Phase Structures of Biopolymer Complexes
Characterisation of Higher Order Biopolymer Structures
"Charge Structures" of ESI– MS of Biopolymers
Ion Mobility– Mass Spectrometry of Polypeptides
HD– Exchange Approaches for Probing Biopolymer Interactions
Chemical Labelling Approaches for Probing Protein Interaction Structures
Characterisation of Biopolymer– Ligand Interactions by Proteolytic Degradation Combined with Mass Spectrometry
Fast Photochemical Oxidation of Proteins (FPOP) and Mass Spectrometry
BIOCHEMICAL APPLICATIONS OF BIOPOLYMER MASS SPECTROMETRY
Introduction
Sample Preparation and Handling in Bioanalytical Applications
Liquid–Liquid Extraction (LLE)
Solid Phase Extraction (SPE)
Solid Phase Microextraction and Supercritical Fluid Extraction (SFE)
Immunoaffinity Extraction (IAE) & Affinity–Isolation
Applications to Primary Structure Analysis of Biopolymers
Protein Sequence Determination
Sequencing and Mutation Analysis in Polynucleotides
Structure Determination of Carbohydrates
Mass Spectrometric Proteome Analysis
Analysis of Post–Translational Protein Structure Modifications
Phosphorylation
Glycosylation
Fatty Acid Acylation
Oxidative Modification of Proteins
Ubiquitin Conjugation
Selected Applications to Analysis of Biopolymer Interactions
Carbohydrate Interactions of Lectins
Structure Analysis of Ion Channels by Chemical Modification and Mass Spectrometry
BIOMEDICAL APPLICATIONS OF BIOPOLYMER MASS SPECTROMETRY
Analysis of Recognition Structures in Molecular Immunology
Identification of Polypeptide Antigen Epitopes
Identification of Antibody Paratopes
Characterization of Oligomerization– Aggregation Pathways in Target Proteins of Neurodegenerative Diseases
Applications to Clinical Diagnostics of Lysosomal Storage Diseases Enzymes
Clinical Applications of Affinity– Mass Spectrometry to Autoimmune Diseases
PERSPECTIVES OF BIOPOLYMER MASS SPECTROMETRY
