The primary structure of human EGF produced by genetic engineering, studied by high-performance tandem mass spectrometry.
The primary structure of human epidermal growth factor (hEGF), which was produced by Escherichia coli using recombinant DNA technique, has been studied by tandem mass spectrometry. The molecular weight of hEGF (about 6200 amu) was determined by fast atom bombardment mass spectrometry. Then reduced and carboxymethylated hEGF was digested by chymotrypsin into seven peptides which could cover the whole sequence of hEGF. The amino acid sequences of five of these seven peptides could be confirmed by tandem mass spectrometry with or without isolation by high-performance liquid chromatography (HPLC). After isolation by HPLC, the other two peptides were digested with trypsin or thermolysin into small peptides, and sequenced by tandem mass spectrometry.
Furuya
M
M
Central Research Laboratories, Ajinomoto Co., Inc., Kawasaki, Japan.
Akashi
S
S
Hirayama
K
K
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D004815:Epidermal Growth Factor; D005818:Genetic Engineering; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins; D013056:Spectrophotometry, Ultraviolet
D011994:Recombinant Proteins; D004815:Epidermal Growth Factor