High-fidelity mass analysis unveils heterogeneity in intact ribosomal particles

Michiel van de Waterbeemd; Kyle L Fort; Dmitriy Boll; Maria Reinhardt-Szyba; Andrew Routh; Alexander Makarov; Albert J R Heck

doi:10.1038/nmeth.4147

High-fidelity mass analysis unveils heterogeneity in intact ribosomal particles

Nat Methods. 2017 Mar;14(3):283-286. doi: 10.1038/nmeth.4147. Epub 2017 Jan 23.

Authors

Michiel van de Waterbeemd^{1

2}, Kyle L Fort^{1

2}, Dmitriy Boll³, Maria Reinhardt-Szyba³, Andrew Routh^{4

5}, Alexander Makarov^{1

3}, Albert J R Heck^{1

2}

Affiliations

¹ Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, the Netherlands.
² Netherlands Proteomics Center, Utrecht, the Netherlands.
³ Thermo Fisher Scientific, Bremen, Germany.
⁴ Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, Texas, USA.
⁵ Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, Texas, USA.

PMID: 28114288
DOI: 10.1038/nmeth.4147

Abstract

Investigation of the structure, assembly and function of protein-nucleic acid macromolecular machines requires multidimensional molecular and structural biology approaches. We describe modifications to an Orbitrap mass spectrometer, enabling high-resolution native MS analysis of 0.8- to 2.3-MDa prokaryotic 30S, 50S and 70S ribosome particles and the 9-MDa Flock House virus. The instrument's improved mass range and sensitivity readily exposes unexpected binding of the ribosome-associated protein SRA.

MeSH terms

Escherichia coli / cytology*
Mass Spectrometry / instrumentation
Mass Spectrometry / methods*
Nodaviridae / genetics
Nodaviridae / ultrastructure*
Protein Binding / physiology
RNA, Long Noncoding / metabolism*
Ribosome Subunits, Large, Bacterial / genetics
Ribosome Subunits, Large, Bacterial / ultrastructure*
Ribosome Subunits, Small, Bacterial / genetics
Ribosome Subunits, Small, Bacterial / ultrastructure*

Substances

RNA, Long Noncoding
steroid receptor RNA activator