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

Journal Article · Sun Apr 01 04:00:00 EDT 1990 · Analytical Chemistry (Washington); (USA)

DOI:https://doi.org/10.1021/ac00206a009· OSTI ID:6279446

Loo, J A; Edmonds, C G; Udseth, H R; Smith, R D ^[1]

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1. Pacific Northwest Laboratory, Richland, WA (USA)

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.

• OSTI ID: 6279446
• Journal Information: Analytical Chemistry (Washington); (USA), Journal Name: Analytical Chemistry (Washington); (USA) Vol. 62:7; ISSN 0003-2700; ISSN ANCHA
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400102* -- Chemical & Spectral Procedures
550200 -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
CARBOXYLIC ACIDS
DATA
DISULFIDES
EXPERIMENTAL DATA
INFORMATION
MASS SPECTROSCOPY
MEASURING INSTRUMENTS
MEASURING METHODS
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PROTEINS
SPECTROSCOPY