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RBS-ID: Comprehensive identification of RNA-binding sites at single amino acid-resolution

ABSTRACT: We developed RBS-ID, which greatly simplifies the RNA moiety by chemical cleavage, reducing the complexity of MS/MS search space to accurately identify and localize RBS in peptides. RBS-ID comprehensively and robustly identifies RNA-binding sites at both proteome and single protein level.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive

ORGANISM(S): Homo Sapiens (human) Escherichia Coli

SUBMITTER: Jong Woo Bae

LAB HEAD: Jong-Seo Kim

PROVIDER: PXD016254 | Pride | 2020-06-02

REPOSITORIES: Pride

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			Action	DRS
	Cas9CBX1_840ug_eUV_RneasyMinelute2vol_HF_HCD_60mG_1.mzXML	Mzxml
	Cas9CBX1_840ug_eUV_RneasyMinelute2vol_HF_HCD_60mG_1.mzid.gz	Mzid
	Cas9CBX1_840ug_eUV_RneasyMinelute2vol_HF_HCD_60mG_1.pride.mgf.gz	Mgf
	Cas9CBX1_840ug_eUV_RneasyMinelute2vol_HF_HCD_60mG_1.pride.mztab.gz	Mztab
	Cas9CBX1_840ug_eUV_RneasyMinelute2vol_HF_HCD_60mG_1.raw	Raw

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Publications

Chemical RNA digestion enables robust RNA-binding site mapping at single amino acid resolution.

Bae Jong Woo JW Kwon S Chul SC Na Yongwoo Y Kim V Narry VN Kim Jong-Seo JS

Nature structural & molecular biology 20200608 7

RNA-binding sites (RBSs) can be identified by liquid chromatography and tandem mass spectrometry analyses of the protein-RNA conjugates created by crosslinking, but RBS mapping remains highly challenging due to the complexity of the formed RNA adducts. Here, we introduce RBS-ID, a method that uses hydrofluoride to fully cleave RNA into mono-nucleosides, thereby minimizing the search space to drastically enhance coverage and to reach single amino acid resolution. Moreover, the simple mono-nucleos ...[more]

PMID: 32514175

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Pytheas SARS-CoV-2 spike protein mRNA sample analysis

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Quantitative photo-crosslinking mass spectrometry reveals protein structure response to environmental changes

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