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Bottom-up and top-down LC-MS/MS of ribosomal purifications

ABSTRACT: Ribosomal purifications of spinach 70S ribosome and human 40S and 60S ribosomal subunits were analyzed with bottom-up LC-MS/MS to identify proteins comprising purified complexes as well as co-purified proteins. In order to assess and quantify proteoforms of ribosomal proteins in ribosomal purifications we employed top-down LC-MS/MS techniques with optimized methods, wherein apart from standard parameters (e.g. collision voltage and dynamic exclusion time) high-resolution and low-resolution were alternately employed in full MS mode.

INSTRUMENT(S): Q Exactive HF

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

SUBMITTER: Sem Tamara

LAB HEAD: Albert J. R. Heck

PROVIDER: PXD008881 | Pride | 2018-06-26

REPOSITORIES: Pride

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Dissecting ribosomal particles throughout the kingdoms of life using advanced hybrid mass spectrometry methods.

van de Waterbeemd Michiel M Tamara Sem S Fort Kyle L KL Damoc Eugen E Franc Vojtech V Bieri Philipp P Itten Martin M Makarov Alexander A Ban Nenad N Heck Albert J R AJR

Nature communications 20180627 1

Biomolecular mass spectrometry has matured strongly over the past decades and has now reached a stage where it can provide deep insights into the structure and composition of large cellular assemblies. Here, we describe a three-tiered hybrid mass spectrometry approach that enables the dissection of macromolecular complexes in order to complement structural studies. To demonstrate the capabilities of the approach, we investigate ribosomes, large ribonucleoprotein particles consisting of a multitu ...[more]

PMID: 29950687

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