2jvr
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Segmental Isotope Labeling of Npl3p
OverviewOverview
The study of multidomain or large proteins in solution by NMR spectroscopy, has been made possible in recent years by the development of new, spectroscopic methods. However, resonance overlap found in large proteins, remains a limiting factor, making resonance assignments and structure, determination of large proteins very difficult. In this study, we present, an expressed protein ligation protocol that can be used for the segmental, isotopic labeling of virtually any multidomain or high molecular mass, protein, independent of both the folding state and the solubility of the, protein fragments, as well as independent of whether the fragments are, interacting. The protocol was applied successfully to two different, multidomain proteins containing RNA recognition motifs (RRMs), heterogeneous nuclear ribonucleoprotein L and Npl3p. High yields of, segmentally labeled proteins could be obtained, allowing characterization, of the interdomain interactions with NMR spectroscopy. We found that the, RRMs of heterogeneous nuclear ribonucleoprotein L interact, whereas those, of Npl3p are independent. Subsequently, the structures of the two RRMs of, Npl3p were determined on the basis of samples in which each RRM was, expressed individually. The two Npl3p RRMs adopt the expected beta alpha, beta beta alpha beta fold.
About this StructureAbout this Structure
2JVR is a Single protein structure of sequence from Saccharomyces cerevisiae. Full crystallographic information is available from OCA.
ReferenceReference
Improved segmental isotope labeling methods for the NMR study of multidomain or large proteins: application to the RRMs of Npl3p and hnRNP L., Skrisovska L, Allain FH, J Mol Biol. 2008 Jan 4;375(1):151-64. Epub 2007 Sep 16. PMID:17936301
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