Characterization of recombinant human epidermal growth factor produced in yeast.
Four different forms of human epidermal growth factor (h-EGF) are found in the culture medium of a recombinant strain of Saccharomyces cerevisiae. These forms were characterized after purification using reverse-phase high-performance liquid chromatography. The most abundant form of secreted recombinant h-EGF has leucine at the carboxyl terminus and is identical with gamma-urogastrone. A second species is identical with the most abundant form except that it lacks the carboxyl-terminal leucine. This form appears to be the product of a carboxypeptidase found in the growth medium. The other two forms of recombinant h-EGF are the respective oxidation products of the above where the single methionine residue has been converted to methionine sulfoxide. These four forms of recombinant h-EGF are fully active; they bind to the EGF receptor of A431 cells as well as stimulate mitotic activity of human foreskin fibroblasts with equal specific activity. The location of the disulfide bonds in the predominant form of recombinant h-EGF was determined following digestion with thermolysin. The amino acid compositions of the resulting peptides showed that the placement of disulfide bonds in recombinant h-EGF is identical with that in murine EGF.
George-Nascimento
C
C
Chiron Research Laboratories, Chiron Corporation, Emeryville, California 94608.
Gyenes
A
A
Halloran
S M
SM
Merryweather
J
J
Valenzuela
P
P
Steimer
K S
KS
Masiarz
F R
FR
Randolph
A
A
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002455:Cell Division / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D004220:Disulfides / Q000032:analysis; D004815:Epidermal Growth Factor / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D008297:Male; D010446:Peptide Fragments / Q000032:analysis; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013820:Thermolysin
D000596:Amino Acids; D004220:Disulfides; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D013820:Thermolysin; D003488:Cyanogen Bromide