Solution structure and dynamics of epidermal growth factor and transforming growth factor alpha.
Circular dichroism (CD) and Fourier transform infrared spectroscopic studies have shown that the secondary structure of transforming growth factor alpha (TGF-alpha) is very similar to that of epidermal growth factor (EGF). The infrared spectra revealed a minor difference between the two proteins, in particular in the beta-sheet structure. A large difference was observed with CD between the two proteins in the apparent conformation each adopts when the disulfide bonds are reduced. Reduced TGF-alpha showed a distinct alpha-helical conformation only at a high trifluoroethanol concentration, whereas reduced EGF assumed an alpha-helical conformation in the absence of trifluoroethanol. This indicates that these two proteins adopt different secondary structures in the absence of disulfide bonds, although they assume similar folding structures in their presence. These data suggest that the disulfide bonds to a large degree dictate the conformation of these two proteins. Additionally, differences in the dynamic behavior between EGF and TGF-alpha were also observed. Infrared experiments showed that the hydrogen-deuterium exchange rate is much higher for TGF-alpha than for EGF, indicating that TGF-alpha is a more flexible molecule. The rate of reduction of the disulfide bonds by dithiothreitol was also faster for TGF-alpha. Therefore, it can be concluded that although EGF and TGF-alpha have a similar overall conformation, TGF-alpha is a more flexible molecule than EGF.
Prestrelski
S J
SJ
Amgen Inc., Amgen Center, Thousand Oaks, California 91320.
Arakawa
T
T
Wu
C S
CS
O'Neal
K D
KD
Westcott
K R
KR
Narhi
L O
LO
D002942:Circular Dichroism; D004815:Epidermal Growth Factor / Q000737:chemistry*; D005583:Fourier Analysis; D006801:Humans; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012996:Solutions; D013055:Spectrophotometry, Infrared; D016211:Transforming Growth Factor alpha / Q000737:chemistry*
D011994:Recombinant Proteins; D012996:Solutions; D016211:Transforming Growth Factor alpha; D004815:Epidermal Growth Factor