Peroxidase-catalyzed bromination of tyrosine, thyroglobulin, and bovine serum albumin: comparison of thyroid peroxidase and lactoperoxidase.
A recent paper (Buchberger, W., 1988, J. Chromatogr. 432, 57) on lactoperoxidase-catalyzed bromination of tyrosine and thyroglobulin stated, without evidence, that thyroid peroxidase (TPO) is able to use bromide as a substrate. This was in disagreement with unpublished experiments previously performed in this laboratory, and we undertook, therefore, to examine this subject further. Highly purified porcine TPO was compared with lactoperoxidase (LPO) and chloroperoxidase (CPO) for ability to catalyze bromination of tyrosine, thyroglobulin, and bovine serum albumin (BSA). The incubation mixture contained 50-100 nM peroxidase, 10-500 microM 82Br-, tyrosine (150 microM), thyroglobulin (0.3 or 1 microM), or BSA (7.5 microM), and a source of H2O2. The latter was either generated by glucose (1 mg/ml)-glucose oxidase (0.5 or 1 micrograms/ml), or added initially as a bolus (100 microM). With TPO, formation of organically bound 82Br was undetectable under all conditions in the pH range 5.4-7.0. Lactoperoxidase and CPO, on the other hand, displayed considerable brominating activity. Lactoperoxidase was much more active at pH 5.4 than at pH 7.0 and was more active with BSA as acceptor than with tyrosine or thyroglobulin. The distribution of 82Br among the various amino acids in LPO-brominated thyroglobulin and BSA was determined by HPLC. As expected, monobromotyrosine and dibromotyrosine together comprised the greatest part of the bound 82Br. However, a surprisingly high percentage (20-25%) was present as monobromohistidine. Evidence was also obtained for the presence of a small percentage of the bound 82Br as tetrabromothyronine. Peroxidase-catalyzed bromination probably depends on the oxidation of Br- to Br+ by the Compound I form of the enzyme. Since oxidation of Br- to Br+ requires a stronger oxidant than oxidation of I- to I+, our results suggest that Compound I of LPO and of CPO has a higher oxidation potential than Compound I of TPO. In vivo experiments with rats on a low iodine diet injected with 82Br- showed that even under conditions of high stimulation by thyrotropic hormone, there is negligible formation of organic bromine in the thyroid. Measurements of thyroid:serum concentration ratios for 82Br- in similar rats provided no evidence that Br- is a substrate for the iodide transport system of the thyroid.
Taurog
A
A
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas 75235-9041.
Dorris
M L
ML
D000818:Animals; D001965:Bromides / Q000378:metabolism* / Q000494:pharmacology; D001967:Bromine Radioisotopes; D002711:Chloride Peroxidase / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006863:Hydrogen-Ion Concentration; D007453:Iodide Peroxidase / Q000378:metabolism*; D007454:Iodides / Q000378:metabolism; D007455:Iodine / Q000378:metabolism; D007784:Lactoperoxidase / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D013552:Swine; D013954:Thyroglobulin / Q000378:metabolism*; D013961:Thyroid Gland / Q000187:drug effects / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
D001965:Bromides; D001967:Bromine Radioisotopes; D007454:Iodides; D012710:Serum Albumin, Bovine; D014443:Tyrosine; D013954:Thyroglobulin; D007455:Iodine; D007784:Lactoperoxidase; D002711:Chloride Peroxidase; D007453:Iodide Peroxidase