Conformational changes in rodent and human alpha-fetoprotein: influence of fatty acids.
Binding, spectral and immunological studies were performed to demonstrate the conformational changes in rodent and human alpha-fetoprotein (AFP) induced by a free fatty acid environment. Scatchard analysis of estradiol (E2) binding to purified rat AFP indicated that unsaturated fatty acids changed the number of binding E2 sites and the apparent E2 equilibrium dissociation constant which varied non-linearly with docosahexaenoic acid concentration. UV spectral analysis of rodent and human AFPs showed that the absorbance minimum of AFP incubated with unsaturated fatty acid (L-AFP) was red-shifted, broadened and less pronounced than that of purified native AFP (N-AFP). Immunochemical studies with specific polyclonal antibodies to purified rodent and human AFPs (N-AFP antibodies) showed that these proteins lost immunoreactivity after incubation with unsaturated fatty acid. N-AFP antibodies recognized fewer epitopes on L-AFP than on N-AFP, whatever the species. Specific anti-rat L-AFP antibodies were used to demonstrate specific epitopes on rat L-AFP. Rat L-AFP antibodies did not recognize rat N-AFP. Saturated fatty acids were without effect on the binding, spectral and immunological properties of rodent and human AFPs. RIA or ELISA values for human AFP from fetal serum, hepatoma serum, and cord serum, were reduced 80, 50 and 5%, respectively, by unsaturated fatty acids. This decrease correlated with the relative percentage of polyunsaturated fatty acid in each biological fluid. Such results indicate that an unsaturated fatty acid environment induces conformational changes in AFP which may modulate the endocrine and immune functions of this protein.
Vallette
G
G
INSERM U.224, Laboratoire de Biochimie, Faculté de Médecine Xavier Bichat, Paris, France.
Vranckx
R
R
Martin
M E
ME
Benassayag
C
C
Nunez
E A
EA
D000818:Animals; D002252:Carbon Tetrachloride Poisoning / Q000097:blood; D002846:Chromatography, Affinity; D004958:Estradiol / Q000378:metabolism; D005230:Fatty Acids, Nonesterified / Q000494:pharmacology*; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D005312:Fetal Blood / Q000032:analysis; D005333:Fetus; D006801:Humans; D007700:Kinetics; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D013045:Species Specificity; D000509:alpha-Fetoproteins / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure
D005230:Fatty Acids, Nonesterified; D005231:Fatty Acids, Unsaturated; D000509:alpha-Fetoproteins; D004958:Estradiol