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CEF research article in F1000 library of top 2% of published articles
February 2011. A CEF research article on the engineering of encodable lanthanide-binding tags into loop regions of proteins has been evaluated by the Faculty of 1000 (F1000) as one of the top 2% of published papers in biology and medicine. The team of authors from Frankfurt, Boston and Cambridge (USA) has shown for the first time that lanthanide-binding peptides can be inserted into the turn connecting two strands of beta-sheets without affecting structure or activity of the protein.
The paper, published on 23 December 2010 by the Journal of the American Chemical Society, shows through a systematic analysis, that Lanthanide-binding tags (LBTs) are versatile tools for the exploitation of the photophysical, paramagnetic, and diffraction properties of lanthanides in NMR and crystallographic structure determination and luminescence applications. LBTs can be attached to proteins via the N-terminus and/or C-terminus, attached to one or two cysteine side chains, or, as shown in the new work, incorporated into loop regions of proteins, relying on well-established molecular biology protocols. For the model protein IL1β, incorporation of the LBT impairs neither the overall fold of the protein nor the binding affinity of Ln3þ to the LBT. As such, the insertion of an LBT can be used for phasing via X-ray crystallography. Furthermore, the loop insertion does not impair binding of IL1β to the soluble domain of the cognate receptor.
Incorporation of LBTs into loop regions of proteins provides a broadly applicable strategy for protein research. Such an improved toolkit is of importance to future applications in phase determination in X-ray crystallography and for the structure determination of large protein complexes by NMR where the LBT can be fused into one NMR-silent binding partner without increased mobility in a similar manner.
Contact
Prof. Dr. Harald Schwalbe
Institut für Organische Chemie und Chemische Biologie
Goethe-Universität Frankfurt
N160/R314
Max-von-Laue-Str. 7
60438 Frankfurt am Main
Telefon: +49 (0)69 798-29737
Fax: +49 (0)69 798-29515
Email: schwalbe@nmr.uni-frankfurt.de
http://schwalbe.org.chemie.uni-frankfurt.de
Link to full paper
Engineering Encodable Lanthanide-Binding Tags into Loop Regions of Proteins.
K Barthelmes, AM Reynolds,..., B Imperiali, H Schwalbe J Am Chem Soc
2010 Dec 23 PMID 21182275 DOI 10.1021/ja104983t
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