Mass Spectrometry of an Intact Virus

Fuerstenau, Stephen D.; Benner, W. Henry; Thomas, John J.; Brugidou, Christophe; Bothner, Brian; Siuzdak, Gary

doi:10.1002/1521-3773(20010202)40:3<541::aid-anie541>3.0.co;2-k

Cited by 157 publications

(43 citation statements)

References 19 publications

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“…This technique has been previously utilized to accurately determine the mass of water droplets, 17 high molecular weight polymers, 18 , 19 nanoparticles, 18 and viruses. 20 , 21 In particular, CD-MS has been recently used to determine the absolute mass of spherical diblock copolymer micelles synthesized via RAFT aqueous emulsion polymerization. 22 , 23 However, as far as we are aware, the present study is the first time that the CD-MS technique has been used to characterize diblock copolymer vesicles.…”

Section: Introductionmentioning

confidence: 99%

Testing the Vesicular Morphology to Destruction: Birth and Death of Diblock Copolymer Vesicles Prepared via Polymerization-Induced Self-Assembly

et al. 2015

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Small angle X-ray scattering (SAXS), electrospray ionization charge detection mass spectrometry (CD-MS), dynamic light scattering (DLS), and transmission electron microscopy (TEM) are used to characterize poly(glycerol monomethacrylate)55-poly(2-hydroxypropyl methacrylate)x (G55-Hx) vesicles prepared by polymerization-induced self-assembly (PISA) using a reversible addition–fragmentation chain transfer (RAFT) aqueous dispersion polymerization formulation. A G55 chain transfer agent is utilized to prepare a series of G55-Hx diblock copolymers, where the mean degree of polymerization (DP) of the membrane-forming block (x) is varied from 200 to 2000. TEM confirms that vesicles with progressively thicker membranes are produced for x = 200–1000, while SAXS indicates a gradual reduction in mean aggregation number for higher x values, which is consistent with CD-MS studies. Both DLS and SAXS studies indicate minimal change in the overall vesicle diameter between x = 400 and 800. Fitting SAXS patterns to a vesicle model enables calculation of the membrane thickness, degree of hydration of the membrane, and the mean vesicle aggregation number. The membrane thickness increases at higher x values, hence the vesicle lumen must become smaller if the external vesicle dimensions remain constant. Geometric considerations indicate that this growth mechanism lowers the total vesicle interfacial area and hence reduces the free energy of the system. However, it also inevitably leads to gradual ingress of the encapsulated water molecules into the vesicle membrane, as confirmed by SAXS analysis. Ultimately, the highly plasticized membranes become insufficiently hydrophobic to stabilize the vesicle morphology when x exceeds 1000, thus this PISA growth mechanism ultimately leads to vesicle “death”.

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Section: Introductionmentioning

confidence: 99%

Testing the Vesicular Morphology to Destruction: Birth and Death of Diblock Copolymer Vesicles Prepared via Polymerization-Induced Self-Assembly

et al. 2015

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“…Another problem is posed by the transient nature of the intermediates formed during assembly and maturation, impeding their purification and analysis. A few decades ago, through pioneering work by the groups of Siuzdak, 5 Robinson, 6 and Heck, 7 native mass spectrometry entered this area of research and tackled several important questions that were less accessible through other techniques. In particular, native MS represents a method that can unambiguously reveal the constituents of a virus, the structural integrity of the particles, and the stoichiometry of the viral structural proteins as well as monitor virus assembly and detect assembly intermediates.…”

Section: Viruses and Virus-like Particlesmentioning

confidence: 99%

Mass Spectrometry-Based Structural Virology

et al. 2020

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“…Eine Kombination aus m/z und Ladungsdetektion in einem ToF-Analysator ergab für den TMV eine Masse von ungefähr 40 MDa mit einer Ungenauigkeit von 15 %. [169] ESI kann daher leicht genutzt werden, um Viren und virale Partikel zu ionisieren. Die akkurate Massenanalyse ist jedoch schwierig, wenn diese Teilchen zu groß oder zu heterogen werden.…”

Section: Massenanalyse Von Intakten Viralen Aggregatenunclassified

Moderne biomolekulare Massenspektrometrie und ihre Bedeutung für die Erforschung der Struktur, der Dynamik und des Aufbaus von Viren

Uetrecht

Heck

2011

Angewandte Chemie

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Entwickelt vor über einem Jahrhundert und beliebter denn je, findet die Massenspektrometrie als analytisches Verfahren heute in beinahe allen Bereichen der Naturwissenschaften Anwendung. Entwicklungen in den letzten beiden Jahrzehnten ermöglichten die Erforschung von Proteinen, sogar intakten Proteinkomplexen, und eine ernsthafte Nutzung in der Strukturbiologie. In diesem Aufsatz beschreiben wir die wachsende Bedeutung der Massenspektrometrie und ihrer technischen Spielarten für die Strukturbiologie anhand einer ausführlichen Betrachtung der strukturellen Virologie. Wir gehen auf die Entwicklung der Massenspektrometrie zu einem Werkzeug ein, das einzigartige Informationen über Struktur, Konformation, Topologie, Aufbau und Funktion von viralen Protein(komplex)en liefern kann, was schließlich in der direkten Analyse von intakten Viruskapsiden mit einer Masse von mehreren Millionen Dalton gipfelt. Die Massenspektrometrie wird heute zur Bearbeitung weitreichender Fragen der Virologie genutzt und hilft so z. B. zu klären, wie Viren aufgebaut werden und wie sie mit ihrem spezifischen Wirt interagieren.

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Mass Spectrometry of an Intact Virus

Cited by 157 publications

References 19 publications

Testing the Vesicular Morphology to Destruction: Birth and Death of Diblock Copolymer Vesicles Prepared via Polymerization-Induced Self-Assembly

Testing the Vesicular Morphology to Destruction: Birth and Death of Diblock Copolymer Vesicles Prepared via Polymerization-Induced Self-Assembly

Mass Spectrometry-Based Structural Virology

Moderne biomolekulare Massenspektrometrie und ihre Bedeutung für die Erforschung der Struktur, der Dynamik und des Aufbaus von Viren

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