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| Subject Categories: RNA | Structural Biology |
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| The EMBO Journal
(2006) 25, 150–162, doi:10.1038/sj.emboj.7600911 Published online 15 December 2005 |
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| Structure of the two most C-terminal RNA recognition motifs of PTB using segmental isotope labeling |
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| Francesca Vitali, Anke Henning, Florian C Oberstrass, Yann Hargous, Sigrid D Auweter, Michèle Erat and Frédéric H-T Allain |
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Institute for Molecular Biology and Biophysics, Swiss Federal Institute of Technology Zurich, ETH-Hönggerberg, Zürich, Switzerland To whom correspondence should be addressed Frédéric H-T Allain, Institute for Molecular Biology and Biophysics, Swiss Federal Institute of Technology Zurich, ETH-Hönggerberg, 8093 Zürich, Switzerland. Tel.: +41 1 633 39 40; Fax: +41 1 633 12 94; E-mail: allain@mol.biol.ethz.ch Received 6 September 2005; Accepted 21 November 2005; Published online 15 December 2005. |
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| Abstract |
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| The polypyrimidine tract binding protein (PTB) is a 58 kDa protein involved in many aspects of RNA metabolism. In this study, we focused our attention on the structure of the two C-terminal RNA recognition motifs (RRM3 and RRM4) of PTB. In a previous study, it was found that the two RRMs are independent in the free state. We recently determined the structure of the same fragment in complex with RNA and found that the two RRMs interact extensively. This difference made us re-evaluate in detail the free protein structure and in particular the interdomain interface. We used a combination of NMR spectroscopy and segmental isotopic labeling to unambiguously study and characterize the interdomain interactions. An improved segmental isotopic labeling protocol was used, enabling us to unambiguously identify 130 interdomain NOEs between the two RRMs and to calculate a very precise structure. The structure reveals a large interdomain interface, resulting in a very unusual positioning of the two RRM domains relative to one another. |
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| Keywords: isotopic labeling, NMR, protein ligation, RRM/RBD/RNP, segmental labeling |
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Top of page MORE ARTICLES LIKE THISThese links to content published by NPG are automatically generated RESEARCHCircumventing tolerance to a human MDM2-derived tumor antigen by TCR gene transfer Nature Immunology Article (01 Oct 2001) The EMBO Journal Article (15 Jun 2000) |
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