Isopycnic centrifugation of thyroid iodoproteins: selectivity of endocytosis.
Isopycnic centrifugation in RbCl was shown to be an effective method both for evaluating the iodine content of thyroglobulin labelled in vivo and for the fractionation of thyroglobulin molecules as a function of their iodine content. Iodination and degradation of thyroid iodoproteins were studied by this method and by zonal centrifugation in sucrose density gradients. Based on these methods it was shown that iodination in vivo is a selective process, 19-S and poorly iodinated thyroglobulin having a higher reactivity toward iodine than 27-S and iodine-rich thyroglobulin. The disappearance of iodoproteins from the thyroid was evaluated by equilibrium, labelling the iodoproteins, blocking iodine incorporation with thiourea derivatives and observing (by sucrose gradient and by RbCl isopycnic centrifugation) at different times the properties of the remaining molecules. Among molecules of different size (19-S and 27-S) and among molecules of the same size (19 S) but different iodine content it was shown that reabsorption from the thyroid gland occurred at the same rate. It was concluded, therefore, that the degradative pathway is essentially a random, non-selective process. Newly iodinated (pulse-labelled) iodoproteins were degraded faster than preexisting molecules. Among the pool of those newly iodinated thyroid proteins, 27-S molecules were reabsorbed faster than 19-S molecules and iodine-rich thyroglobulin molecules were reabsorbed faster than the iodine-poor ones. Since iodination in vivo occurs as repeated pulses of iodine incorporation, it is suggested that this latter phenomenon is a regulatory mechanism which minimizes degradation of molecules which are iodine-poor and have a lower hormonal content.
D000818:Animals; D002499:Centrifugation, Density Gradient; D004705:Endocytosis; D007467:Iodoproteins / Q000378:metabolism*; D008297:Male; D009929:Organ Size; D051381:Rats; D013954:Thyroglobulin / Q000302:isolation & purification / Q000378:metabolism; D013961:Thyroid Gland / Q000378:metabolism*
D007467:Iodoproteins; D013954:Thyroglobulin