Structural characterization and biological activity of recombinant human epidermal growth factor proteins with different N-terminal sequences.
The primary structures and molecular homogeneity of recombinant human epidermal growth factors from different suppliers were characterized and their biological activities evaluated by a standard DNA synthesis assay. Molecular weight determinations using 252Cf-plasma-desorption and electrospray mass spectrometry in combination with N- and C-terminal sequence analysis and determination of intramolecular disulfide bridges revealed that one recombinant protein had the correct human-identical structure (54 aa residues; 6347 Da). In contrast, a second recombinant protein (7020 Da) was found to contain a pentapeptide (KKYPR) insert following its N-terminal methionine. This structural variant showed a significant reduction in its capacity to stimulate DNA synthesis.
Svoboda
M
M
Faculty of Chemistry, University of Konstanz, Germany.
Bauhofer
A
A
Schwind
P
P
Bade
E
E
Rasched
I
I
Przybylski
M
M
D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000096:biosynthesis; D004815:Epidermal Growth Factor / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000032:analysis
D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D004247:DNA