Cu(I) Disrupts the Structure and Function of the Nonclassical Zinc Finger Protein Tristetraprolin (TTP)
| العنوان: | Cu(I) Disrupts the Structure and Function of the Nonclassical Zinc Finger Protein Tristetraprolin (TTP) |
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| المؤلفون: | Geoffrey D. Shimberg, Sarah L. J. Michel, Kathryn E. Splan, Heather M. Neu, Kiwon Ok |
| المصدر: | Inorganic Chemistry. 56:6838-6848 |
| بيانات النشر: | American Chemical Society (ACS), 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Protein Conformation, Tristetraprolin, chemistry.chemical_element, Inflammation, Peptide, Zinc, 010402 general chemistry, 01 natural sciences, Article, Inorganic Chemistry, 03 medical and health sciences, hemic and lymphatic diseases, medicine, heterocyclic compounds, Physical and Theoretical Chemistry, neoplasms, chemistry.chemical_classification, Zinc finger, Chemistry, RNA, respiratory system, 0104 chemical sciences, Structure and function, Cell biology, 030104 developmental biology, Biochemistry, Mrna level, medicine.symptom, therapeutics, Copper, Protein Binding |
| الوصف: | Tristetraprolin (TTP) is a nonclassical zinc finger (ZF) protein that plays a key role in regulating inflammatory response. TTP regulates cytokines at the mRNA level by binding to AU-rich sequences present at the 3′-untranslated region, forming a complex that is then degraded. TTP contains two conserved CCCH domains with the sequence Cys-X8CysX5CysX3His that are activated to bind RNA when zinc is coordinated. During inflammation, copper levels are elevated, which is associated with increased inflammatory response. A potential target for Cu(I) during inflammation is TTP. To determine whether Cu(I) binds to TTP and how Cu(I) can affect TTP/RNA binding, two TTP constructs were prepared. One construct contained just the first CCCH domain (TTP-1D) and serves as a peptide model for a CCCH domain; the second construct contains both CCCH domains (TTP-2D) and is functional (binds RNA) when Zn(II) is coordinated. Cu(I) binding to TTP-1D was assessed via electronic absorption spectroscopy titrations, and Cu(I) binding to TTP-2D was assessed via both absorption spectroscopy and a spin filter/inductively coupled plasma mass spectrometry (ICP-MS) assay. Cu(I) binds to TTP-1D with a 1:1 stoichiometry and to TTP-2D with a 3:1 stoichiometry. The CD spectrum of Cu(I)-TTP-2D did not exhibit any secondary structure, matching that of apo-TTP-2D, while Zn(II)-TTP-2D exhibited a secondary structure. Measurement of RNA binding via fluorescence anisotropy revealed that Cu(I)-TTP-2D does not bind to the TTP-2D RNA target sequence UUUAUUUAUUU with any measurable affinity, while Zn(II)-TTP-2D binds to this site with nanomolar affinity. Similarly, addition of Cu(I) to the Zn(II)-TTP-2D/RNA complex resulted in inhibition of RNA binding. Together, these data indicate that, while Cu(I) binds to TTP-2D, it does not result in a folded or functional protein and that Cu(I) inhibits Zn(II)-TTP-2D/RNA binding. |
| تدمد: | 1520-510X 0020-1669 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::65cabaf8526f84acc47f2445a38f5d6f https://doi.org/10.1021/acs.inorgchem.7b00125 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....65cabaf8526f84acc47f2445a38f5d6f |
| قاعدة البيانات: | OpenAIRE |
Find this issue from the American Chemical Society | تدمد: | 1520510X 00201669 |
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