2JVO

Segmental isotope labeling of Npl3

2JVO の概要

• エントリーDOI: 10.2210/pdb2jvo/pdb
• 関連するPDBエントリー: 2JVR
• NMR情報: BMRB: 15485
• 分子名称: Nucleolar protein 3 (1 entity in total)
• 機能のキーワード: protein, nucleus, phosphorylation, ribonucleoprotein, ribosome biogenesis, rna-binding, rrna processing, rna binding protein
• 由来する生物種: Saccharomyces cerevisiae (baker's yeast)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 12292.84

構造登録者

Skrisovska, L.,Allain, F.H.-T. (登録日: 2007-09-24, 公開日: 2007-12-18, 最終更新日: 2024-05-01)

主引用文献

Skrisovska, L.,Allain, F.H.
Improved segmental isotope labeling methods for the NMR study of multidomain or large proteins: application to the RRMs of Npl3p and hnRNP L
J.Mol.Biol., 375:151-164, 2008

PubMed Abstract: 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.

PubMed: 17936301
DOI: 10.1016/j.jmb.2007.09.030

実験手法

SOLUTION NMR