Protein Ladder Sequencing

A new approach to protein sequencing is described. It consists of two steps: (i) laddergenerating chemistry, the controlled generation from a polypeptide chain by wet chemistry of a family of sequence-defining peptide fragments, each differing from the next by one amino acid; and (ii) data readout,...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 1993-10, Vol.262 (5130), p.89-92
Main Authors: Chait, Brian T., Wang, Rong, Beavis, Ronald C., Stephen B. H. Kent
Format: Article
Language:English
Subjects:
Accuracy
Amino Acid Sequence
Amino acid sequencing
Amino acids
Analytical, structural and metabolic biochemistry
Average linear density
Biochemistry
Biological and medical sciences
Chemical mixtures
Chemistry
Datasets
Fibrinopeptide B - chemistry
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Ladders
Mass Spectrometry
Mass spectroscopy
Measurement
Molecular Sequence Data
Organic Chemistry
Peptides - chemistry
Polypeptides
Post translational modification
Protein engineering
Proteins
Sequence analysis
Sequence Analysis - methods
Sequencing
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Summary:A new approach to protein sequencing is described. It consists of two steps: (i) laddergenerating chemistry, the controlled generation from a polypeptide chain by wet chemistry of a family of sequence-defining peptide fragments, each differing from the next by one amino acid; and (ii) data readout, a one-step readout of the resulting protein sequencing ladder by matrix-assisted laser-desorption mass spectrometry. Each amino acid was identified from the mass difference between successive peaks, and the position in the data set defined the sequence of the original peptide chain. This method was used to directly locate a phosphoserine residue in a phosphopeptide. The protein ladder sequencing method lends itself to very high sample throughput at very low per cycle cost.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.8211132