Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry

Summary: Standard methods for mass analysis measure ensembles of thousand to millions of molecules. This approach enables analysis of monodisperse recombinant proteins, whereas some heterogeneous protein assemblies pose a significant challenge, whereby co-occurring stoichiometries, sub-complexes, an...

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Autores principales:	Szu-Hsueh Lai, Sem Tamara, Albert J.R. Heck
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Physical chemistry Biochemistry Structural biology Science Q
Acceso en línea:	https://doaj.org/article/2b24d1a3985d4ec0966e7f67b9c53aee
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spelling	oai:doaj.org-article:2b24d1a3985d4ec0966e7f67b9c53aee2021-11-20T05:08:20ZSingle-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry2589-004210.1016/j.isci.2021.103211https://doaj.org/article/2b24d1a3985d4ec0966e7f67b9c53aee2021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2589004221011792https://doaj.org/toc/2589-0042Summary: Standard methods for mass analysis measure ensembles of thousand to millions of molecules. This approach enables analysis of monodisperse recombinant proteins, whereas some heterogeneous protein assemblies pose a significant challenge, whereby co-occurring stoichiometries, sub-complexes, and modifications hamper analysis using native mass spectrometry. To tackle the challenges posed by mass heterogeneity, single-particle methods may come to the rescue. Recently, two such approaches have been introduced, namely, mass photometry (MP) and Orbitrap-based charge detection mass spectrometry (CDMS). Both methods assess masses of individual molecules, albeit adhering to distinct physical principles. To evaluate these methods side by side, we analyzed a set of ribosomal particles, representing polydisperse ribonucleoprotein assemblies in the MDa range. MP and CDMS provide accurate masses for intact ribosomes and enable quantitative analysis of concomitant distinct particles within each ribosome sample. Here, we discuss pros and cons of these single-molecule techniques, also in the context of other techniques used for mass analysis.Szu-Hsueh LaiSem TamaraAlbert J.R. HeckElsevierarticlePhysical chemistryBiochemistryStructural biologyScienceQENiScience, Vol 24, Iss 11, Pp 103211- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Physical chemistry Biochemistry Structural biology Science Q
spellingShingle	Physical chemistry Biochemistry Structural biology Science Q Szu-Hsueh Lai Sem Tamara Albert J.R. Heck Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry
description	Summary: Standard methods for mass analysis measure ensembles of thousand to millions of molecules. This approach enables analysis of monodisperse recombinant proteins, whereas some heterogeneous protein assemblies pose a significant challenge, whereby co-occurring stoichiometries, sub-complexes, and modifications hamper analysis using native mass spectrometry. To tackle the challenges posed by mass heterogeneity, single-particle methods may come to the rescue. Recently, two such approaches have been introduced, namely, mass photometry (MP) and Orbitrap-based charge detection mass spectrometry (CDMS). Both methods assess masses of individual molecules, albeit adhering to distinct physical principles. To evaluate these methods side by side, we analyzed a set of ribosomal particles, representing polydisperse ribonucleoprotein assemblies in the MDa range. MP and CDMS provide accurate masses for intact ribosomes and enable quantitative analysis of concomitant distinct particles within each ribosome sample. Here, we discuss pros and cons of these single-molecule techniques, also in the context of other techniques used for mass analysis.
format	article
author	Szu-Hsueh Lai Sem Tamara Albert J.R. Heck
author_facet	Szu-Hsueh Lai Sem Tamara Albert J.R. Heck
author_sort	Szu-Hsueh Lai
title	Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry
title_short	Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry
title_full	Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry
title_fullStr	Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry
title_full_unstemmed	Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry
title_sort	single-particle mass analysis of intact ribosomes by mass photometry and orbitrap-based charge detection mass spectrometry
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/2b24d1a3985d4ec0966e7f67b9c53aee
work_keys_str_mv	AT szuhsuehlai singleparticlemassanalysisofintactribosomesbymassphotometryandorbitrapbasedchargedetectionmassspectrometry AT semtamara singleparticlemassanalysisofintactribosomesbymassphotometryandorbitrapbasedchargedetectionmassspectrometry AT albertjrheck singleparticlemassanalysisofintactribosomesbymassphotometryandorbitrapbasedchargedetectionmassspectrometry
_version_	1718419532195823616

Single-particle mass analysis of intact ribosomes by mass photometry and Orbitrap-based charge detection mass spectrometry

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