Quantitation of Thyroid Hormone Binding to Anti-Thyroxine Antibody Fab Fragment by Native Mass Spectrometry
| Autor: | Janne Jänis, Senthil K. Thangaraj, Henri O. Arola, Juha Rouvinen, Antti Tullila, Tarja K. Nevanen |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
General Chemical Engineering
chemistry.chemical_element Iodine 01 natural sciences Article 03 medical and health sciences chemistry.chemical_compound medicine Thyroid hormone binding QD1-999 030304 developmental biology 0303 health sciences biology 010401 analytical chemistry Thyroid General Chemistry Ligand (biochemistry) Affinities 0104 chemical sciences Chemistry medicine.anatomical_structure chemistry Biochemistry Thyronine biology.protein Antibody Hormone |
| Zdroj: | ACS Omega Thangaraj, S K, Arola, H, Tullila, A, Nevanen, T K, Rouvinen, J & Jänis, J 2019, ' Quantitation of Thyroid Hormone Binding to Anti-Thyroxine Antibody Fab Fragment by Native Mass Spectrometry ', ACS Omega, vol. 4, no. 20, pp. 18718-18724 . https://doi.org/10.1021/acsomega.9b02659 ACS Omega, Vol 4, Iss 20, Pp 18718-18724 (2019) |
| ISSN: | 2470-1343 |
| DOI: | 10.1021/acsomega.9b02659 |
| Popis: | Thyroid hormones are important regulatory hormones, acting on nearly every cell in the body. The two main thyroid hormones are l-thyroxine (tetraiodo-l-thyronine, T4) and 3,3′,5-triiodo-l-thyronine (T3), which are produced in the thyroid gland and secreted into the blood stream. Other important thyroid hormone metabolites are 3,3′-diiodo-l-thyronine (T2) and l-thyronine (T0), which may show increased levels in circulation due to dietary iodine deficiency or other medical disorders. Owing to their central role in cellular functions, sensitive and specific detection methods for thyroid hormones are needed. In this work, native mass spectrometry (MS) was used to quantitate thyroid hormone binding to the anti-T4 antibody Fab fragment. First, the binding affinity for T2 was determined via direct ligand titration experiments. Then, the affinities for the other ligands were determined by competition experiments using T2 as the "low-affinity" reference ligand. The highest affinity was measured for T3, followed by T4, T2, and T0 (Kd = 29, 3.4, and 260 nM and 130 μM, respectively). Thus, it is evident that the number and positions of the iodine substituents within the thyronine rings are important for the ligand binding affinity of anti-T4 Fab. Surprisingly, structurally related tetrahalogen bisphenols were also able to bind to anti-T4 Fab with nanomolar affinities. |
| Databáze: | OpenAIRE |
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