Characterization and primary structures of bovine and porcine thyroxine-binding globulin.
Thyroxine-binding globulin (TBG) is the major serum transport protein for iodothyronines in most of the large, omni- or herbivorous mammals. Characterization of human TBG (hTBG), including its 20 known natural variants, allowed the identification of the ligand-binding site and a correlation of diminished synthesis or loss of function with mutations in the TBG gene. Further refinement of the structure-function correlation, especially the high binding affinity and heat stability, requires characterization of other mammalian TBGs, of which only rat and sheep TBG were available. We now present some of the chemical and physical properties of bovine TBG (bTBG) and porcine TBG (pTBG) and their primary structures deduced from their cDNA sequences. The serum concentrations of bTBG and pTBG estimated by Scatchard analysis of T(4)-binding were similar to hTBG. The T(4)-binding affinity of human, bovine and porcine TBGs were all similar, at 1.2x10(10) M(-1). However, heat stability of the animal TBGs was reduced, with a half life of denaturation of 7 min (bTBG) and 5 min (pTBG) at 55 degreeC, compared with 21 min for hTBG. Nucleotide alignment revealed identity with hTBG of 85.5% (bTBG) and 83.7% (pTBG) and amino acid identity of 82.8% (bTBG) and 82.6% (pTBG). As expected, the relevant parts of the ligand-binding domain (amino acids 215-291, and 363-395) were highly conserved at more than 95% similarity. Comparison of the five known mammalian TBGs allows focusing of future mutagenesis experiments to further characterize the properties of the molecule.
Janssen
Onno E
OE
Department of Medicine, Division of Endocrinology, Universitatsklinikum Essen, Hufelandstrasse 55, D-45122, Essen, Germany. onno.janssen@uni-essen.de
Lahner
Harald
H
Grasberger
Helmut
H
Spring
Sabine A
SA
Saller
Bernhard
B
Mann
Klaus
K
Refetoff
Samuel
S
Einspanier
Ralf
R
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D018076:DNA, Complementary / Q000737:chemistry; D019143:Evolution, Molecular; D005091:Exons; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D013552:Swine; D013963:Thyroid Hormones / Q000097:blood; D013975:Thyroxine-Binding Proteins / Q000032:analysis / Q000737:chemistry* / Q000235:genetics
D000596:Amino Acids; D018076:DNA, Complementary; D013963:Thyroid Hormones; D013975:Thyroxine-Binding Proteins