Mass Spectrometry of Proteins Directly from Polyacrylamide Gels
and
Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109-0674, and Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, Michigan 48105
Received for review December 19, 1995. Accepted March
4, 1996.
Abstract:
The direct combination of thin-layer gel
electrophoresis
and matrix-assisted laser desorption/ionization mass
spectrometry has been demonstrated with good sensitivity
and mass accuracy, offering potential advantages in speed
and reduced complexity. Mass spectra have been obtained from isoelectric focusing, sodium dodecyl sulfate,
and native gels with as little as 660 fmol of 
- and
-chain
bovine hemoglobin and 1 pmol of horse heart myoglobin
loaded. CNBr digests were performed in situ, and the
products were probed in-gel. Noncovalent complexes
such as multimeric protein systems, enzyme inhibitor
complexes, and protein-ligand complexes can also be
characterized when gel electrophoresis is run under
nondenaturing conditions. This approach shows promise
for simplifying the interface between gel electrophoresis
and mass spectrometry.
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