Protein sequencing by tandem mass spectrometry. |
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| Authors: | D F Hunt J R Yates rd J Shabanowitz S Winston C R Hauer |
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| Affiliation: | D F Hunt, J R Yates, 3rd, J Shabanowitz, S Winston, and C R Hauer |
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| Abstract: | Methodology for determining amino acid sequences of proteins by tandem mass spectrometry is described. The approach involves enzymatic and/or chemical degradation of the protein to a collection of peptides which are then fractionated by high-performance liquid chromatography. Each fraction, containing as many as 10-15 peptides, is then analyzed directly, without further purification, by a combination of liquid secondary-ion/collision-activated dissociation mass spectrometry on a multianalyzer instrument. Interpretation of collision-activated dissociation mass spectra is described, and results are presented from a study of soluble peptides produced by treatment of apolipoprotein B with cyanogen bromide and trypsin. |
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