Thyroxine binding by human transthyretin variants: mutations at position 119, but not position 54, increase thyroxine binding affinity.
A mutation at codon 119 in the transthyretin (TTR) gene leads to a substitution of methionine for threonine at this position in the circulating protein. As the amino acid at position 119 is located in the T4 binding channel, mutations here may affect the binding of T4 by TTR. A previous study has shown an increase in the amount of hormone carried by the TTRMet119 variant. To determine whether this increase in binding was due to a change in affinity or capacity, TTR was partially purified from normal individuals and those with the TTRMet119 mutation. The isolation procedure was a rapid, single step passage through Blue Sepharose. With normal serum, the resulting protein bound T4 with a single site of intermediate affinity (Ka, 1.63 +/- 0.36 x 10(7) L/mol). No sites of higher or lower affinity were detected. Comparisons of binding capacity and immunoreactive TTR concentrations showed that the preparations bound T4 with a molar ratio between 1-2. With TTRMet119 serum, the T4 affinity was approximately doubled [Ka, 3.40 +/- 0.76 x 10(7) L/mol (+/- SD); P < 0.001] with no change in binding capacity. This doubling in affinity explains the observed T4 levels of about 120 nmol/L in individuals with this mutation. Binding of rT3 to TTRMet119 was increased approximately 5-fold over normal. Identical experiments with TTRGly54, in which glycine is substituted for glutamine, showed that the T4 affinity of this variant was unchanged from normal. These results suggest that the TTRMet119 mutation leads to secretion of a normal concentration of TTR that has a raised affinity for T4. Depending on their location, mutations in the TTR gene may lead to an increase or no change in T4 binding by the secreted protein.
Curtis
A J
AJ
Ewen Downie Metabolic Unit, Alfred Hospital, Melbourne, Victoria, Australia.
Scrimshaw
B J
BJ
Topliss
D J
DJ
Stockigt
J R
JR
George
P M
PM
Barlow
J W
JW
D014644:Genetic Variation; D005971:Glutamates / Q000032:analysis; D018698:Glutamic Acid; D005998:Glycine / Q000032:analysis; D006801:Humans; D007223:Infant; D008715:Methionine / Q000032:analysis; D008722:Methods; D009154:Mutation / Q000235:genetics; D011228:Prealbumin / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D013912:Threonine / Q000032:analysis; D013974:Thyroxine / Q000032:analysis / Q000378:metabolism*
D005971:Glutamates; D011228:Prealbumin; D013912:Threonine; D018698:Glutamic Acid; D008715:Methionine; D013974:Thyroxine; D005998:Glycine