Binding of thyroxine to pig transthyretin, its cDNA structure, and other properties.
Thyroxine binding to proteins in pig plasma during electrophoresis was observed in the albumin, but not in the prealbumin and post-albumin regions. Transthyretin could be identified in medium from in vitro pig choroid plexus incubations by size and number of subunits and a very high rate of synthesis and secretion. Its electrophoretic mobility was intermediate between that of thyroxine-binding globulin and albumin. It bound thyroxine, retinol-binding protein, anti-(rat transthyretin) antibodies and behaved similarly to transthyretins from other vertebrate species when plasma was extracted with phenol. Inhibition experiments with the synthetic flavonoid F 21388, analysing the binding of thyroxine, suggested that transthyretin is not a major thyroxine carrier in the bloodstream of pigs. Cloning and sequencing of transthyretin cDNA from both choroid plexus and liver showed that the same transthyretin mRNA is expressed in pig choroid plexus and liver. The amino acid sequence derived from the nucleotide sequence revealed that pig transthyretin differs from the transthyretins of all other studied vertebrate species by an unusual C-terminal extension consisting of the amino acids glycine, alanine and leucine. This extension results from the mutation of a stop codon into a codon for glycine. The unusual C-terminal extensions do not seem to interfere with the access of thyroxine to its binding site in the central channel of transthyretin.
Duan
W
W
Russell Grimwade School of Biochemistry, University of Melbourne, Australia.
Richardson
S J
SJ
Köhrle
J
J
Chang
L
L
Southwell
B R
BR
Harms
P J
PJ
Brack
C M
CM
Pettersson
T M
TM
Schreiber
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002831:Choroid Plexus / Q000378:metabolism; D002846:Chromatography, Affinity; D018076:DNA, Complementary / Q000737:chemistry*; D008969:Molecular Sequence Data; D011228:Prealbumin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine; D013974:Thyroxine / Q000378:metabolism*
D018076:DNA, Complementary; D011228:Prealbumin; D011994:Recombinant Proteins; D013974:Thyroxine