Hormone-containing peptides from normal and goiter human thyroglobulins.
A series of low iodine human thyroglobulin samples derived from colloid-rich goiter tissue was examined by HPLC mapping of tryptic digests and compared to normal human thyroglobulin. These samples ranged in iodine content from 2 to 8 gram-atoms of iodine (g.a. I) per mole and were not further iodinated in vitro. Peptides containing the principal hormonogenic sequence were detected using the long wavelength absorbance of the iodotyrosine derivatives at 325 nm. Two such peptides were isolated and sequenced. Their thyroxine content was confirmed by radioimmunoassay. The number of 325-nm-absorbing peaks was significantly lower in the normally iodinated human thyroglobulin than that observed the thyroglobulins of cattle and dog. This suggests a more restricted iodination in the human protein. Sodium dodecyl sulfate gel patterns of the reduced and alkylated proteins showed significant molecular size heterogeneity in all of the samples. Polypeptide fragments ranged in molecular size from approximately 330 to 45 kDa in the goiter derived material and from approximately 330 to 15 kDa in the normal human material. This difference between the proteins is consistent with earlier observations that peptides less than 45 kDa appear concomitantly with hormone formation. These data confirm that the human thyroglobulin molecule is capable of forming at least limited amounts of thyroid hormone at iodine levels as low as 4 g.a. I per mole. The hormone detected in this study was located at residue 5 near the amino terminus of the thyroglobulin molecule.
Dziadik-Turner
C
C
Department of Biochemistry, University of Kansas Medical Center, Kansas City 66103.
Hamilton
J W
JW
Taurog
A
A
Rawitch
A B
AB
D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006042:Goiter / Q000378:metabolism*; D006801:Humans; D007455:Iodine / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D010455:Peptides / Q000032:analysis*; D011506:Proteins / Q000032:analysis; D011863:Radioimmunoassay; D013954:Thyroglobulin / Q000032:analysis*; D013963:Thyroid Hormones / Q000032:analysis*; D013974:Thyroxine / Q000032:analysis; D014284:Triiodothyronine / Q000032:analysis
D010446:Peptide Fragments; D010455:Peptides; D011506:Proteins; D013963:Thyroid Hormones; D014284:Triiodothyronine; D013954:Thyroglobulin; D007455:Iodine; D013974:Thyroxine