Structure of the two most C-terminal RNA recognition motifs of PTB using segmental isotope labeling

被引：78

作者：

Vitali, F ^{[1
]}

Henning, A ^{[1
]}

Oberstrass, FC ^{[1
]}

Hargous, Y ^{[1
]}

Auweter, SD ^{[1
]}

Erat, M ^{[1
]}

Allain, FHT ^{[1
]}

机构：

[1] ETH Honggerberg, Inst Mol Biol & Biophys, Swiss Fed Inst Technol, CH-8093 Zurich, Switzerland

来源：

EMBO JOURNAL | 2006年 / 25卷 / 01期

关键词：

isotopic labeling; NMR; protein ligation; RRM/RBD/RNP; segmental labeling;

D O I：

10.1038/sj.emboj.7600911

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：150 / 162

页数：13

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