19F labelling of 14-3-3ζ recombinant protein for 19F NMR spectroscopy
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14740%2F21%3A00122345" target="_blank" >RIV/00216224:14740/21:00122345 - isvavai.cz</a>
Result on the web
<a href="http://www.csbmb.cz" target="_blank" >http://www.csbmb.cz</a>
DOI - Digital Object Identifier
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Alternative languages
Result language
angličtina
Original language name
19F labelling of 14-3-3ζ recombinant protein for 19F NMR spectroscopy
Original language description
<sup>19</sup>F NMR has been a very useful complementary approach to traditional techniques - double labelling by <sup>13</sup>C and <sup>15</sup>N - due to the excellent magnetic properties of the <sup>19</sup>F isotope. 1) It has a spin ½ and strong dipolar coupling (useful in nuclear Overhauser effect spectroscopy), 2) High sensitivity (83% relative to <sup>1</sup>H) and broad chemical shift range (up to 400 ppm), 3) <sup>19</sup>F is 100% abundant in nature and virtually non‑present in biologically relevant samples. Selective <sup>19</sup>F isotopic labelling is therefore an outstanding technique to monitor region‑specific changes in protein structure owing to minimal background signal. Here, we present our progress in the preparation of 14-3-3 protein samples labelled with <sup>19</sup>F-modified aromatic amino acids (AAs): 5-<sup>19</sup>F-Trp, 4-<sup>19</sup>F-Phe, and 3-<sup>19</sup>F-Tyr. Even though we used identical protocols, different AAs had different incorporation efficiency rates. <sup>19</sup>F tryptophan was readily incorporated with 100% efficiency. However, the extent of incorporation of <sup>19</sup>F phenylalanine and tyrosine ranged only between 30-50%, presumably due to the similar biosynthetic pathways or suboptimal culture conditions. On the other hand, the amount and purity of samples was sufficient for pilot titration experiments, as demonstrated by well-resolved 1D <sup>19</sup>F NMR spectra.
Czech name
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Czech description
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Classification
Type
O - Miscellaneous
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
<a href="/en/project/LTAUSA18168" target="_blank" >LTAUSA18168: Selective NMR labelling as a tool for characterization of protein complexes involved in neurodegenerative diseases.</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2021
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů