دورية أكاديمية
Chemical tools for study of phosphohistidine: generation of selective Τ‐phosphohistidine and Π‐phosphohistidine antibodies
| العنوان: | Chemical tools for study of phosphohistidine: generation of selective Τ‐phosphohistidine and Π‐phosphohistidine antibodies |
|---|---|
| المؤلفون: | Makwana, M., dos Santos Souza, C., Pickup, B.T., Thompson,, M.J., Lomada, S.K., Feng, Y., Wieland, T., Jackson, R.F.W., Muimo, R. |
| بيانات النشر: | Wiley |
| سنة النشر: | 2023 |
| المجموعة: | White Rose Research Online (Universities of Leeds, Sheffield & York) |
| الوصف: | Non-hydrolysable stable analogues of τ-phosphohistidine (τ-pHis) and π-pHis have been designed aided by electrostatic surface potential calculations, and subsequently synthesized. The τ-pHis and π-pHis analogues (phosphopyrazole 8 and pyridyl amino amide 13, respectively) were used as haptens to generate pHis polyclonal antibodies. Both τ-pHis and π-pHis conjugates in the form of a BSA-glutaraldehyde-τ-pHis and BSA-glutaraldehyde-π-pHis were synthesized and characterized by 31P NMR spectroscopy. Commercially available τ-pHis (SC56-2) and π-pHis (SC1-1; SC50-3) monoclonal antibodies were used to show that the BSA-G-τ-pHis and BSA-G-π-pHis conjugates could be used to assess the selectivity of pHis antibodies in a competitive ELISA. Subsequently, the selectivity of the generated pHis antibodies generated using phosphopyrazole 8 and pyridyl amino amide 13 as haptens was assessed by competitive ELISA against His, pSer, pThr, pTyr, τ-pHis and π-pHis. Antibodies generated using the phosphopyrazole 8 as a hapten were found to be selective for τ-pHis, and antibodies generated using the pyridyl amino amide 13 were found to be selective for π-pHis. Both τ- and π-pHis antibodies were shown to be effective in immunological experiments, including ELISA, western blot, and immunofluorescence. The τ-pHis antibody was also shown to be useful in the immunoprecipitation of proteins containing pHis. |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | text |
| اللغة: | English |
| العلاقة: | https://eprints.whiterose.ac.uk/201847/1/ChemBioChem%20-%202023%20-%20Makwana%20-%20Chemical%20Tools%20for%20Studying%20Phosphohistidine%20%20Generation%20of%20Selective%20%20%E2%80%90Phosphohistidine%20and.pdfTest; Makwana, M., dos Santos Souza, C. orcid.org/0000-0001-5314-6263 , Pickup, B.T. orcid.org/0000-0002-2728-5486 et al. (6 more authors) (2023) Chemical tools for study of phosphohistidine: generation of selective Τ‐phosphohistidine and Π‐phosphohistidine antibodies. ChemBioChem. e202300182. ISSN 1439-4227 |
| الإتاحة: | https://eprints.whiterose.ac.uk/201847Test/ https://eprints.whiterose.ac.uk/201847/1/ChemBioChem%20-%202023%20-%20Makwana%20-%20Chemical%20Tools%20for%20Studying%20Phosphohistidine%20%20Generation%20of%20Selective%20%20%E2%80%90Phosphohistidine%20and.pdfTest |
| حقوق: | cc_by_nc_nd_4 |
| رقم الانضمام: | edsbas.EE518638 |
| قاعدة البيانات: | BASE |
| الوصف غير متاح. |