Effect of reducing disulfide-containing proteins on electrospray ionization mass spectra

Electrospray ionization produces multiply charged molecular ions for biomolecules with molecular weights in excess of 100,000. This allows mass spectrometers with limited mass-to-charge range to extend their molecular weight range by a factor equal to the number of charges. The maximum number of obs...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 1990-04, Vol.62 (7), p.693-698
Main Authors: Loo, Joseph A, Edmonds, Charles G, Udseth, Harold R, Smith, Richard D
Format: Article
Language:English
Subjects:
400102 - Chemical & Spectral Procedures
550200 - Biochemistry
AMINO ACIDS
Analytical, structural and metabolic biochemistry
BASIC BIOLOGICAL SCIENCES
Biological and medical sciences
CARBOXYLIC ACIDS
DATA
DISULFIDES
Dithiothreitol
Electrochemistry
EXPERIMENTAL DATA
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
INFORMATION
INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY
Mass Spectrometry - methods
MASS SPECTROSCOPY
MEASURING INSTRUMENTS
MEASURING METHODS
Molecular Weight
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
Oxidation-Reduction
Protein Conformation
PROTEINS
Proteins - analysis
SPECTROSCOPY
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Description
Summary:Electrospray ionization produces multiply charged molecular ions for biomolecules with molecular weights in excess of 100,000. This allows mass spectrometers with limited mass-to-charge range to extend their molecular weight range by a factor equal to the number of charges. The maximum number of observed charges for peptides and smaller proteins correlates well with the number of basic amino acid residues (Arg, Lys, His), except for disulfide-containing molecules, such as lysozyme and bovine albumin. However, reduction of disulfide linkages with 1,4-dithiothreitol (Cleland's reagent) may allow the protein to be in an extended conformation and make "buried" basic residues available for protonation to yield higher charged molecular ions by the electrospray ionization process. For larger proteins reduction of disulfide bridges greatly increases the maximum charge state, but charging of basic amino acid residues remains less efficient than for smaller proteins.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac00206a009