The dissociation of b-TSH. The effect of sodium dodecyl sulfate, pH, urea and temperature.

In the present study SDS polyacrylamide gel electrophoresis was used to reinvestigate the conditions under which b-TSH dissociates into its constituent subunits. The effects of SDS, itself, urea, acid pH and temperature were assessed by the decrease in the amount of undissociated TSH and the increase in the dissociated components as revealed by SDS gel electrophoresis. In SDS alone 25--30% dissociation occurred. Pre-treatment with increasing concentrations of urea up to 7.1 M increased the degree of dissociation to 70% after h at room temperature. Maximum dissociation was achieved by treatment at pH 2.5 and 37 degree C for 1 h. In 1 M propionic acid approximately 10% of the TSH was still undissociated after 18 h at 25 degree C.

Bioaffinity chromatography of thyrotropin using immobilized concanavalin A.

The behavior of crude preparations of whale and bovine thyrotropins was studied on an affinity column packed with concanavalin A-Sepharose. Only a small portion of the proteins applied was adsorbed to the column and eluted quantitatively with 0.5 M methyl-alpha-D-glucoside or -mannoside. Immunoreactive as well as hormonally active thyrotropin was recovered exclusively in the absorbed fraction. The usefulness of this chromatographic procedure for the group separation of pituitary glycoprotein hormones is discussed.

Estimation of the secretion rate of thyrotropin in man.

The plasma concentration of a pituitary hormone is determined by the rate of secretion, degradation, and the volume of distribution of that hormone. Using a radioimmunoassay for human thyrotropin (TSH) and human TSH-(131)I, we have estimated the rates of degradation and distribution of TSH in man and calculated the rate of secretion. Either 0.5 or 5 mug of TSH-(131)I with specific activities of 1 to 50 muc per mug was administered intravenously to 12 euthyroid subjects. Serial determinations were made of TSH-(131)I, and the half-time of disappearance (t((1/2))) was thus estimated. The average t((1/2)) in euthyroid subjects was 53.9 minutes with a volume of distribution averaging 5.8% of body weight. The mean endogenous plasma TSH concentration was 1.8 mmug per ml (2.7 muU per ml in terms of the human TSH reference standard A). The mean total TSH pool, excluding the pituitary, was 5.8 mug (8.7 mU). From these data the mean secretion rate of TSH in euthyroid man was calculated to be 110.1 mug per day (165.2 mU). Similar data were estimated for 3 mildly hypothyroid patients. The t((1/2)) were 75.1, 97.1, and 83.6 minutes, with a mean of 85.3 minutes (1.6 times normal). The mean TSH pool was 58.1 mug (10 times normal). The secretion rate was 688.7 mug per day (1,033.1 mU). In other hypothyroid patients, plasma TSH levels ranging from 6 to 230 mmug per ml (9 to 345 muU) have been found. If similar half-times and a normal distribution volume are assumed, the secretion rate of TSH in hypothyroid patients can be estimated to range from about 260 to 15,350 mug per day (390 to 23,025 mU) or from about 2 to 307 times normal. Therefore, the elevated plasma TSH levels found in hypothyroidism are a result of both slower degradation and increase in rate of secretion.

Thyrocalcitonin: ultracentrifugation in gradients of sucrose.

Analysis of thyrocalcitonin, by density-gradient ultracentrifugation at high speed, showed that its molecular weight (5000 to 6000) is considerably less than was heretofore recognized.