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利用分段同位素标记RNA的交联和串联质谱对蛋白质-RNA复合物进行结构建模。

Structural modeling of protein-RNA complexes using crosslinking of segmentally isotope-labeled RNA and MS/MS.

作者信息

Dorn G, Leitner A, Boudet J, Campagne S, von Schroetter C, Moursy A, Aebersold R, Allain F H-T

机构信息

Department of Biology, Institute for Molecular Biology and Biophysics, ETH Zürich, 8093 Zürich, Switzerland.

Department of Biology, Institute of Molecular Systems Biology, ETH Zürich, 8093 Zürich, Switzerland.

出版信息

Nat Methods. 2017 May;14(5):487-490. doi: 10.1038/nmeth.4235. Epub 2017 Mar 27.

DOI:10.1038/nmeth.4235
PMID:28346450
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5505470/
Abstract

Ribonucleoproteins (RNPs) are key regulators of cellular function. We established an efficient approach, crosslinking of segmentally isotope-labeled RNA and tandem mass spectrometry (CLIR-MS/MS), to localize protein-RNA interactions simultaneously at amino acid and nucleotide resolution. The approach was tested on polypyrimidine tract binding protein 1 and U1 small nuclear RNP. Our method provides distance restraints to support integrative atomic-scale structural modeling and to gain mechanistic insights into RNP-regulated processes.

摘要

核糖核蛋白（RNP）是细胞功能的关键调节因子。我们建立了一种高效的方法，即分段同位素标记RNA交联与串联质谱分析（CLIR-MS/MS），以在氨基酸和核苷酸分辨率水平上同时定位蛋白质-RNA相互作用。该方法已在多嘧啶序列结合蛋白1和U1小核核糖核蛋白上进行了测试。我们的方法提供了距离限制，以支持整合的原子尺度结构建模，并深入了解RNP调节的过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/8ebdbcfe614f/emss-71740-f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/4bdca5a9f9f4/emss-71740-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/384b1f8b1f5f/emss-71740-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/8ebdbcfe614f/emss-71740-f003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/4bdca5a9f9f4/emss-71740-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/384b1f8b1f5f/emss-71740-f002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa6d/5505470/8ebdbcfe614f/emss-71740-f003.jpg

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