New Methods for the Study of Biomolecular Complexes

A NATO Advanced Research Workshop entitled New Methods for the Study of Molecular Aggregates was held at Tbe Lodge at Kananaskis Village, Alberta, Canada from 16 -20 June 1996. In fact the meeting was entirely concerned with the problem of analyzing biomolecular complexes, so the title of these proceedings has been altered to give a more precise description of the content. Tbe workshop was hosted by the time-of-flight group of the Department of Physics at the University of Manitoba, and was attended by 64 participants from around the world. '!\venty-one invited talks were given and 27 papers were presented as posters. Of the 48 contributions, 22 papers (12 orals, 10 posters) are included in these proceedings. Tbe subject of the conference was the investigation of noncovalent biomolecular complexes, with particular focus on the application of mass spectrometry to their characterization. '!\vo new ionization techniques introduced in the late 1980s, electrospray ionization (ES I) and matrix-assisted laser desorptionlionization (MALDI), resulted in a breakthrough in mass spectrometry, enabling its use in molecular weight and primary structure determination of biopolymers larger than 100 kDa. Recently it has been discovered that ESI mass spectrometry mayaiso be used to characterize complexes containing noncovalent interactions, thus opening new perspectives for supramolecular chemistry. ESI mass spectrometry has the advantage that the sampie is introduced from a homogenous solution which can be maintained at near physiological conditions of pR, concentration, and temperature.

Klappentext

The particular focus of the present book is the application of mass spectrometry to the investigation and characterization of noncovalent biomolecular complexes. Both electrospray ionization and matrix-assisted laser desorption/ionization methods are discussed. Electrospray ionization mass spectrometry is particularly effective for studying noncovalent interactions because the sample is introduced from an homogeneous solution which can be maintained under near physiological conditions of pH, concentration and temperature. Mass spectra of large complexes (several hundred kDa) have been obtained using sector, quadrupole, time-of-flight and ion cyclotron resonance mass spectrometry. The use of other techniques for analyzing supramolecular complexes is also presented, particularly in combination with mass spectrometry. These include such emerging techniques as surface plasmon resonance as well as more established methods, such as X-ray crystallography and NMR.

Inhalt
A General Approach to Deciphering how Proteins Work Using Synthetic Chemistry and Mass Spectrometry.- Approaches to the Characterisation of Tertiary and Supramolecular Protein Structures by Combination of Protein Chemistry and Mass Spectrometry.- Protein-Ligand Binding Studied by Amide Hydrogen Exchange and Mass Spectrometry.- Structural Studies of Protein-Protein Interactions Using Proteolytic Footprinting and MALDI/MS.- Investigation of the Effect of Protein Oxidation upon the Lipid-Binding Activity of the Cerebroside-Sulfate Activator Protein (Saposin B).- The Use of Chemical Ligation for Investigation of the Structure of Duplexes Formed by Linear and Circular Oligonucleotides.- Studying Noncovalent Protein-RNA Interactions and Drug Binding by Electrospray Ionization Mass Spectrometry.- Measurement of Noncovalent Complexes with High m/z by Electrospray Time-of-Flight Mass Spectrometry.- Non-Covalent Complexes of Oligonucleotides Observed Using Electrospray Ionization Mass Spectrometry.- Observations of Non-Covalent Complexes of Citrate Synthase and NADH by ESI/TOF Mass Spectrometry.- Gas-Phase Dissociation Energies of Hydrogen-Bonded Complexes in a Quadrupole Ion Trap Mass Spectrometer.- Electrospray Ionization FTICR Spectrometry of Metalloproteins.- On the Determination of the Ion Internal Energy in an Electrospray Source.- Observation of Protein Heptamer Formation for Chemically Ligated Early Pregnancy Factor.- Matrix-Assisted Laser Desorption Ionization of Non-Covalent Complexes.- Characterization of Tertiary Structure States and Specific Noncovalent Complexes of Proteins by UV-Matrix-Assisted Laser-Desorption/Ionization Mass Spectrometry.- Non-Covalent Hetero- and Homo-Oligomeric Protein Complexes Reassociate Differentially during MALDI-MS Analysis.- The Detection of Non-Covalent Interactions in Nucleic Acids Using MALDI.- Mass Spectrometric Methods for Biomolecular Characterization.- X-Ray Crystallographic and Mass Spectrometric Structure Determination and Functional Characterisation of Succinylated Porin from Rhodobacter Capsulatus: Implications for Ion Selectivity and Single-Channel Conductance.- Structural Relation of Peridinin-Chlorophyll A-Protein (PCP) and Proteins with Globin-like Fold A Testable Hypothesis for its Folding and Assembly Pathway.- The Development of NMR Methods to Study Protein Structure and Dynamics.- BIAcore Technology A Marriage Partner to Mass Spectrometry?.- Biospecific Interactions Measured by Capillary Electrophoresis.- Application of Neutron Scattering Techniques to Problems of Biological Relevance.- List of Participants.