European Journal of Biochemistry · 1967 · 2.8K citations · 9 references
Life SciencesBiochemistryMolecular BiologyProtein BiochemistryProteomicsSequence AnalysisProtein ChemistryProtein DegradationProtein SeparationProtein Sequence AnalysisProtein StructureAmino Acid AnalysisAmino Acid Composition AnalysisAmino Acid Degradation PathwaysAmino Acid SequencingAmino AcidsAutomated Thin-layer Chromatography
The protein sequenator automatically determines amino acid sequences in proteins and peptides. It operates via a phenylisothiocyanate degradation scheme that forms a phenylthiocarbamyl derivative, releases the N‑terminal amino acid as thiazolinone, converts thiazolinones to phenylthiohydantoins for thin‑layer chromatography, and requires about 0.25 µmoles of protein. The process achieves 15.4 cycles per 24 h with >98 % yield per cycle and successfully sequenced the first 60 N‑terminal amino acids of apomyoglobin from humpback whale.
The protein sequenator is an instrument for the automatic determination of amino acid sequences in proteins and peptides. It operates on the principle of the phenylisothiocyanate degradation scheme. The automated process embraces the formation of the phenylthiocarbamyl derivative of the protein and the splitting off of the N‐terminal amino acid as thiazolinone. The degradation proceeds at a rate of 15.4 cycles in 24 hours and with a yield in the individual cycle in excess of 98%. The material requirements are approximately 0.25 μmoles of protein. The thiazolinones are converted to the corresponding phenylthiohydantoins in a separate operation, and the latter identified by thin layer chromatography. The process has been applied to the whole molecule of apomyoglobin from the humpback whale, and it has been possible to establish the sequence of the first 60 amino acids from the N‐terminal end.
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Amino-Acid Sequence of Sperm Whale Myoglobin
Allen B. Edmundson · Nature · 1965 · 399 citations
Life Sciences, Biochemistry, Zoology +13
On the Mechanism of the Phenyl Isothiocyanate Degradation of Peptides.
Pehr Edman, Bengt Lindberg, H. I. Waterman et al. · Acta chemica Scandinavica/Acta chemica Scandinavica. B, Organic chemistry and biochemistry/Acta chemica Scandinavica. A, Physical and inorganic chemistry/Acta chemica Scandinavica. Series B. Organic chemistry and biochemistry/Acta chemica Scandinavica. Series A, Physical and inorganic chemistry · 1956 · 321 citations · Full text
Identification and Semiquantitative Determination of Phenyl Thiohydantoins.
Pehr Edman, John Sjöquist, K. Lunde et al. · Acta chemica Scandinavica/Acta chemica Scandinavica. B, Organic chemistry and biochemistry/Acta chemica Scandinavica. A, Physical and inorganic chemistry/Acta chemica Scandinavica. Series B. Organic chemistry and biochemistry/Acta chemica Scandinavica. Series A, Physical and inorganic chemistry · 1956 · 304 citations · Full text