Determination of proteolytic hydrolysis of thyroglobulin
| Autor: | Eugène H.J.M. Jansen, F.X. Rolaf van Leeuwen, Lambert Doorn |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Bromides
Male Thyroid Hormones medicine.medical_treatment Proteolysis Pronase Thyroglobulin chemistry.chemical_compound Endopeptidases medicine Animals Chromatography High Pressure Liquid Chromatography Chymotrypsin Diiodotyrosine biology medicine.diagnostic_test Hydrolysis Sodium Proteolytic enzymes General Chemistry Monoiodotyrosine Sodium Compounds Rats chemistry Biochemistry Thyronine biology.protein Cattle |
| Zdroj: | Journal of chromatography. 566(2) |
| Popis: | A method for the determination of the free thyronine- and tyrosine-like amino acids in the thyroidal protein thyroglobulin is presented. The compounds of interest are monoiodotyrosine, diiodotyrosine, thyronine, diiodothyronine, triidothyronine and tetraiodothyronine. The extent of proteolysis was followed by high-performance liquid chromatographic monitoring of both the remaining peptides and the formation of the free thyroidal amino acids. Total hydrolysis was achieved by a combination of proteolytic enzymes. A number of enzymes were tested, such as trypsin, chymotrypsin, pronase, aminopeptidase-M, carboxypeptidase-A, carboxypeptidase-P and carboxypeptidase-Y. The best combination turned out to be pronase followed by aminopeptidase-M. The relative amounts of the enzymes, with respect to the substrate thyroglobulin, and the time of incubation were optimized to achieve total proteolysis in 4 h. The method was applied successfully to samples from a toxicological experiment with sodium bromide. |
| Databáze: | OpenAIRE |
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