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      Native Mass Spectrometry: What is in the Name?

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          Abstract

          Electrospray ionization mass spectrometry (ESI-MS) is nowadays one of the cornerstones of biomolecular mass spectrometry and proteomics. Advances in sample preparation and mass analyzers have enabled researchers to extract much more information from biological samples than just the molecular weight. In particular, relevant for structural biology, noncovalent protein–protein and protein–ligand complexes can now also be analyzed by MS. For these types of analyses, assemblies need to be retained in their native quaternary state in the gas phase. This initial small niche of biomolecular mass spectrometry, nowadays often referred to as “native MS,” has come to maturation over the last two decades, with dozens of laboratories using it to study mostly protein assemblies, but also DNA and RNA-protein assemblies, with the goal to define structure–function relationships. In this perspective, we describe the origins of and (re)define the term native MS, portraying in detail what we meant by “native MS,” when the term was coined and also describing what it does (according to us) not entail. Additionally, we describe a few examples highlighting what native MS is, showing its successes to date while illustrating the wide scope this technology has in solving complex biological questions.

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          A generic protein purification method for protein complex characterization and proteome exploration.

          We have developed a generic procedure to purify proteins expressed at their natural level under native conditions using a novel tandem affinity purification (TAP) tag. The TAP tag allows the rapid purification of complexes from a relatively small number of cells without prior knowledge of the complex composition, activity, or function. Combined with mass spectrometry, the TAP strategy allows for the identification of proteins interacting with a given target protein. The TAP method has been tested in yeast but should be applicable to other cells or organisms.
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            Protein and polymer analyses up tom/z 100 000 by laser ionization time-of-flight mass spectrometry

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              Analytical properties of the nanoelectrospray ion source.

              The nanoelectrospray ion source (nanoES) has recently been developed and described theoretically. It is different from conventional electrospray sources and from other miniaturized electrospray sources by (i) its 1-2 microns spraying orifice achieved by pulling the spraying capillary to a fine tip, (ii) its very low flow rate of approximately 20 nL/min and the small size of droplets it generates, and (iii) the absence of solvent pumps and inlet valves. The fabrication and operation of nanoES needles is described in detail. Solutions with up to 0.1 M salt contents could be sprayed without sheath flow or pneumatic assist. Improved desolvation in nanoES led to instrument-limited resolution of the signals of a glycoprotein and the ability to signal average extensively allowed the C-terminal sequencing of a 40 kDa protein. Extensive mass spectrometric and tandem mass spectrometric investigation of the components of an unseparated peptide mixture was demonstrated by verification of 93% of the sequence of carbonic anhydrase. A rapid and robust desalting/concentration step coupled to the nanoES procedure allows the direct analysis of impure samples such as peptide mixtures extracted after in-gel digestion.
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                Author and article information

                Contributors
                a.j.r.heck@uu.nl
                Journal
                J Am Soc Mass Spectrom
                J. Am. Soc. Mass Spectrom
                Journal of the American Society for Mass Spectrometry
                Springer US (New York )
                1044-0305
                1879-1123
                1 December 2016
                1 December 2016
                2017
                : 28
                : 1
                : 5-13
                Affiliations
                [1 ]Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584CH Utrecht, The Netherlands
                [2 ]Netherlands Proteomics Center, Padualaan 8, 3584CH Utrecht, The Netherlands
                Article
                1545
                10.1007/s13361-016-1545-3
                5174146
                27909974
                f74e53ff-99e1-45a4-b416-fb46292835db
                © The Author(s) 2016

                Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

                History
                : 18 October 2016
                : 25 October 2016
                : 28 October 2016
                Funding
                Funded by: Utrecht University
                Categories
                Focus: 31st Asilomar Conference, Native MS-based Structural Biology: Account & Perspective
                Custom metadata
                © American Society for Mass Spectrometry 2017

                Analytical chemistry
                electrospray ionization mass spectrometry (esi-ms),native mass spectrometry

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