Book contents
- Frontmatter
- Contents
- Foreword by D. M. Engelman
- Foreword by Pierre Joliot
- Preface
- Introduction: Molecular biophysics at the beginning of the twenty-first century: from ensemble measurements to single-molecule detection
- Part A Biological macromolecules and physical tools
- Part B Mass spectrometry
- Chapter B1 Mass and charge
- Chapter B2 Structure function studies
- Part C Thermodynamics
- Part D Hydrodynamics
- Part E Optical spectroscopy
- Part F Optical microscopy
- Part G X-ray and neutron diffraction
- Part H Electron diffraction
- Part I Molecular dynamics
- Part J Nuclear magnetic resonance
- References
- Index of eminent scientists
- Subject Index
- References
Chapter B2 - Structure function studies
from Part B - Mass spectrometry
Published online by Cambridge University Press: 05 November 2012
Book contents
- Frontmatter
- Contents
- Foreword by D. M. Engelman
- Foreword by Pierre Joliot
- Preface
- Introduction: Molecular biophysics at the beginning of the twenty-first century: from ensemble measurements to single-molecule detection
- Part A Biological macromolecules and physical tools
- Part B Mass spectrometry
- Chapter B1 Mass and charge
- Chapter B2 Structure function studies
- Part C Thermodynamics
- Part D Hydrodynamics
- Part E Optical spectroscopy
- Part F Optical microscopy
- Part G X-ray and neutron diffraction
- Part H Electron diffraction
- Part I Molecular dynamics
- Part J Nuclear magnetic resonance
- References
- Index of eminent scientists
- Subject Index
- References
Summary
Protein structure and function
ESI and MALDI have become increasingly useful for the mass spectrometric analysis of proteins. The two ionisation techniques have been exploited to study protein folding, to characterise non-covalent protein complexes, to map protein function and for many other applications (Comment B2.1).
Comment B2.1: Mass spectrometry and X-ray crystallography
It is very useful to use mass spectrometry prior to X-ray crystallography. Indeed, the determination of a protein structure at atomic resolution takes a considerable investment of time and effort (see Chapter G3). Mass spectrometry permits a rapid check of the correctness of the accepted primary structure of a protein, and high-resolution determination of the purity of protein preparations. The information is particularly important for proteins obtained by recombinant techniques, which are subject to a number a special sources of error, including unanticipated mutations, modifications, termination and proteolitic degradation. A simple molecular mass measurement is so informative and time saving that there is a reason to obtain the mass spectrum of virtually every protein before it is subject to X-ray crystallography. It is especially important for proteins produced with special amino acid residues (e.g. selenomethionine), for NMR with 13C and/or 15N enrichment and for neutron small-angle scattering with specific deuteration. In the last case the incorporation of deuterium into biological macromolecules by biosynthetic methods is routinely determined by mass spectrometry measurements.
Mass determination
ESI and MALDI have made the mass analysis of proteins a routine procedure.
- Type
- Chapter
- Information
- Methods in Molecular BiophysicsStructure, Dynamics, Function, pp. 136 - 170Publisher: Cambridge University PressPrint publication year: 2007
References
(1994). Mass spectrometry – useful tool for protein X-ray crystallographer and NMR spectroscopist. Structure, 2, 465–467.CrossRefGoogle ScholarPubMed
, and (1997). Mass spectrometry as a readout of protein structure and function. Mass Spectr. Rev., 16, 165–179.3.0.CO;2-F>CrossRefGoogle ScholarPubMed
, and (1999). Disassembly of protein complexes in the gas phase. Curr. Opin. Struct. Biol., 9, 135–141.CrossRefGoogle ScholarPubMed
(2000). Protein complexes and analysis of their assembly by mass spectrometry. Curr. Opin. Struct. Biol., 10, 601–606.CrossRefGoogle ScholarPubMed
, and (2001). Detection of multiple protein conformational ensembles in solution via deconvolution of charge-state distribution in ESI MS. Anal. Chem., 73, 4763–4772.CrossRefGoogle Scholar
, , , , and (2002). Peptide end sequencing by orthogonal MALDI tandem mass spectroscopy. J. Proteome Res., 1, 63–71.CrossRefGoogle Scholar
, , et al. (1999). Direct analysis of protein complexes using mass spectrometry. Nature Biotechnol., 17, 676–682.CrossRefGoogle ScholarPubMed
, and (1997). Protein identification from 2-DE gels by MALDI mass spectrometry.Mass Spectr. Rev., 16, 145–162.3.0.CO;2-H>CrossRefGoogle ScholarPubMed
, and (2001). Perspectives for mass spectrometry and functional proteomics. Mass Spectr. Rev., 20, 1–57.3.0.CO;2-J>CrossRefGoogle Scholar
, , et al. (2001). Virtual 2-D gel electrophoresis: visualization and analysis of the E. coli proteome by mass spectrometry. Anal. Chem., 73, 4063–4070.CrossRefGoogle Scholar
, , and (1996). Mass spectrometry of nucleic acids. Mass Spectr. Rev., 15, 67–138.3.0.CO;2-8>CrossRefGoogle ScholarPubMed
(1996). DNA sequencing by mass spectrometry. J. Mass Spectr., 31, 1203–1215.3.0.CO;2-3>CrossRefGoogle ScholarPubMed
, , and (1998). Infrared MALDI mass spectrometry of large nucleic acids. Science, 281, 260–262.CrossRefGoogle ScholarPubMed
, and (1998). Application of mass spectrometry to the characterization of oligonucleotides and nucleic acids. Curr. Opin. Biotechnol., 9, 25–34.CrossRefGoogle Scholar
(1989). Animal glycosphingolipids as membrane attachment sites for bacteria. Ann. Rev. Biochem., 58, 309–350.CrossRefGoogle ScholarPubMed
, , et al. (1998). Electrospray ionization and matrix-assisted laser desorption/ionisation Fourier transform ion cyclotron resonance mass spectrometry of permethylated oligosaccarides. Anal. Chem., 70, 857–864.CrossRefGoogle Scholar
, , et al. (2000). Detection and selective dissociation of intact ribosomes in a mass spectrometer. Proc. Natl. Acad. Sci. USA, 97, 5185–5190.CrossRefGoogle Scholar
(2001). MALDI-TOF mass spectrometry of bacteria. Mass Spectr. Rev., 20, 172–194.CrossRefGoogle Scholar
, and (1999). Imaging with mass spectrometry. Chem. Rev., 99, 2977–3005.CrossRefGoogle ScholarPubMed
(1994). Mass spectrometry – useful tool for protein X-ray crystallographer and NMR spectroscopist. Structure, 2, 465–467.CrossRefGoogle ScholarPubMed
, and (1997). Mass spectrometry as a readout of protein structure and function. Mass Spectr. Rev., 16, 165–179.3.0.CO;2-F>CrossRefGoogle ScholarPubMed
, and (1999). Disassembly of protein complexes in the gas phase. Curr. Opin. Struct. Biol., 9, 135–141.CrossRefGoogle ScholarPubMed
(2000). Protein complexes and analysis of their assembly by mass spectrometry. Curr. Opin. Struct. Biol., 10, 601–606.CrossRefGoogle ScholarPubMed
, and (2001). Detection of multiple protein conformational ensembles in solution via deconvolution of charge-state distribution in ESI MS. Anal. Chem., 73, 4763–4772.CrossRefGoogle Scholar
, , , , and (2002). Peptide end sequencing by orthogonal MALDI tandem mass spectroscopy. J. Proteome Res., 1, 63–71.CrossRefGoogle Scholar
, , et al. (1999). Direct analysis of protein complexes using mass spectrometry. Nature Biotechnol., 17, 676–682.CrossRefGoogle ScholarPubMed
, and (1997). Protein identification from 2-DE gels by MALDI mass spectrometry.Mass Spectr. Rev., 16, 145–162.3.0.CO;2-H>CrossRefGoogle ScholarPubMed
, and (2001). Perspectives for mass spectrometry and functional proteomics. Mass Spectr. Rev., 20, 1–57.3.0.CO;2-J>CrossRefGoogle Scholar
, , et al. (2001). Virtual 2-D gel electrophoresis: visualization and analysis of the E. coli proteome by mass spectrometry. Anal. Chem., 73, 4063–4070.CrossRefGoogle Scholar
, , and (1996). Mass spectrometry of nucleic acids. Mass Spectr. Rev., 15, 67–138.3.0.CO;2-8>CrossRefGoogle ScholarPubMed
(1996). DNA sequencing by mass spectrometry. J. Mass Spectr., 31, 1203–1215.3.0.CO;2-3>CrossRefGoogle ScholarPubMed
, , and (1998). Infrared MALDI mass spectrometry of large nucleic acids. Science, 281, 260–262.CrossRefGoogle ScholarPubMed
, and (1998). Application of mass spectrometry to the characterization of oligonucleotides and nucleic acids. Curr. Opin. Biotechnol., 9, 25–34.CrossRefGoogle Scholar
(1989). Animal glycosphingolipids as membrane attachment sites for bacteria. Ann. Rev. Biochem., 58, 309–350.CrossRefGoogle ScholarPubMed
, , et al. (1998). Electrospray ionization and matrix-assisted laser desorption/ionisation Fourier transform ion cyclotron resonance mass spectrometry of permethylated oligosaccarides. Anal. Chem., 70, 857–864.CrossRefGoogle Scholar
, , et al. (2000). Detection and selective dissociation of intact ribosomes in a mass spectrometer. Proc. Natl. Acad. Sci. USA, 97, 5185–5190.CrossRefGoogle Scholar
(2001). MALDI-TOF mass spectrometry of bacteria. Mass Spectr. Rev., 20, 172–194.CrossRefGoogle Scholar
, and (1999). Imaging with mass spectrometry. Chem. Rev., 99, 2977–3005.CrossRefGoogle ScholarPubMed