Mass spectrometry and viral analysis

Siuzdak, Gary; Bothner, Brian; Yeager, Mark; Brugidou, Christophe; Fauquet, Claude; Hoey, Kenway; Change, Cheng-Ming

doi:10.1016/s1074-5521(96)90083-6

Cited by 188 publications

(169 citation statements)

References 18 publications

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“…This experiment demonstrates that singly charged bioparticles as viruses can be size-separated and collected by means of the PDMA system and used for further investigations. This proof-of-principle experiment corroborates the experiments of Siuzdak et al, demonstrating the viability of TMV and rice yellow mottle virus after ESI and passage of a quadrupole analyzer (vacuum system) with subsequent soft collection by means of a gelatinous layer [35]. It will open up new avenues of research (in terms micropreparative collection) in a number of fields, where the integrity and viability of nanobioparticles are of importance (e.g., drug carrier or vaccine particles).…”

Section: Collection Of Intact Tobacco Mosaic Virussupporting

confidence: 88%

Nano ES GEMMA and PDMA, new tools for the analysis of nanobioparticles—Protein complexes, lipoparticles, and viruses

Allmaier

Laschober

Szymanski

2008

J. Am. Soc. Mass Spectrom.

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Differential mobility analysis (DMA) is a technique suited for size analysis as well as preparative collection of airborne nanosized airborne particles. In the recent decade, the analysis of intact viruses, proteins, DNA fragments, polymers, and inorganic nanoparticles was possible when combining this method with a nano-electrospray charge-reduction source for producing aerosols from a sample solution/suspensions. Mass analysis of high molecular weight noncovalent complexes is also possible with this methodology due to the linear correlation of the electrophoretic mobility diameter and the molecular mass. In this work, we present the analysis (size and molecular mass) of high molecular weight multimers (noncovalent functional homocomplex) of Jack bean urease in a mass range from 275 kDa up to 2.5 MDa, with mainly present tri-and hexamers but also higher oligomers of the 91 kDa monomer subunit. In a second experiment, the size analysis of intact very-low-density (ϳ35 nm), low-density (ϳ22 nm) and high-density lipoparticles (ϳ10 nm), which are heterocomplexes consisting of cholesterol, lipids, and proteins in different ratios, is presented. Results from mobility analysis were in excellent agreement with particle diameters found in literature. The last presented experiment demonstrates size analysis of a rod-like virus and selective sampling of a selected size fraction of electrosprayed, singly-charged tobacco mosaic virus particles. Sampling and subsequent transmission electron microscopic investigations of a specific size fraction (40 nm electrophoretic mobility diameter) revealed the folding of virus particles during the electrospray and charge reduction (electrical stress) as well as solvent evaporation (mechanical stress) process, leading to an observed geometry of 150 (length) ϫ 35 (width) nm (average cylindrical geometry of unsprayed intact virus 300 ϫ 18 nm). and viruses. Analysis of the formation process, stoichiometry, and molecular weight of those complexes can be very challenging as noncovalent interactions can easily be destroyed or biased when leaving specific native conditions [5], and as their molecular mass can easily exceed the working range of modern mass spectrometry. For these reasons, a demand for new analytical techniques, which preserve noncovalent interactions, deliver molecular mass and/or size related information (in contrast to capillary electrophoresis and native gel electrophoresis) and which have a working range that exceed conventional mass spectrometry, exists.Differential mobility analysis (DMA) is a technique developed to classify charged aerosolized particles under ambient pressure according to their electrophoretic mobility diameter [6]. In the recent decade, its working range was extended from m down to the nm size range [7], or in terms of molecular mass, into the kDa to GDa molecular mass range, thus closing the gap between classic aerosol particle technology (low m into mm range) and mass spectrometry (sub-nm to 10 nm). The size range of 10 to 200 nm is therefore of great in...

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Section: Collection Of Intact Tobacco Mosaic Virussupporting

confidence: 88%

Nano ES GEMMA and PDMA, new tools for the analysis of nanobioparticles—Protein complexes, lipoparticles, and viruses

Allmaier

Laschober

Szymanski

2008

J. Am. Soc. Mass Spectrom.

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“…Furthermore, the gentle nature of the transfer of proteins from the electrospray droplet to the gas phase means that perturbation of the solution structure of a protein is minimal (39), and there is now significant evidence to suggest that the correlation between the solution and gas phase properties of a protein is significant (27,40,41). An impressive and increasing number of ESI-MS-based approaches now exist with regard to the characterization of the conformational dynamics of proteins (38).…”

Section: Discussionmentioning

confidence: 99%

Co-populated Conformational Ensembles of β2-Microglobulin Uncovered Quantitatively by Electrospray Ionization Mass Spectrometry

Borysik

Radford²,

Ashcroft

2004

Journal of Biological Chemistry

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Ordered assembly of monomeric human ␤ 2 -microglobulin (␤ 2 m) into amyloid fibrils is associated with the disorder hemodialysis-related amyloidosis. Previously, we have shown that under acidic conditions (pH <5.0 at 37°C), wild-type ␤ 2 m assembles spontaneously into fibrils with different morphologies. Under these conditions, ␤ 2 m populates a number of different conformational states in vitro. However, this equilibrium mixture of conformationally different species is difficult to resolve using ensemble techniques such as nuclear magnetic resonance or circular dichroism. Here we use electrospray ionization mass spectrometry to resolve different species of ␤ 2 m populated between pH 6.0 and 2.0. We show that by linear deconvolution of the charge state distributions, the extent to which each conformational ensemble is populated throughout the pH range can be determined and quantified. Thus, at pH 3.6, conditions under which short fibrils are produced, the conformational ensemble is dominated by a charge state distribution centered on the 9؉ ions. By contrast, under more acidic conditions (pH 2.6), where long straight fibrils are formed, the charge state distribution is dominated by the 10؉ and 11؉ ions. The data are reinforced by investigations on two variants of ␤ 2 m (V9A and F30A) that have reduced stability to pH denaturation and show changes in the pH dependence of the charge state distribution that correlate with the decrease in stability measured by tryptophan fluorescence. The data highlight the potential of electrospray ionization mass spectrometry to resolve and quantify complex mixtures of different conformational species, one or more of which may be important in the formation of amyloid.␤ 2 -microglobulin (␤ 2 m) 1 is one of a number of proteins known to aggregate into insoluble amyloid deposits in vivo (1-3). These amyloid deposits have been linked to various human maladies such as Alzheimer's disease (4), Huntington's chorea (5), and senile systemic amyloidosis (6). However, for the majority of the ϳ20 known amyloid-related proteins, the structural mechanism of amyloid formation is still unclear, and, for several proteins, the role that different aggregated states play in the manifestation of the disease symptoms remains unresolved (7).␤ 2 m is a small (11,860-dalton) extracellular protein that forms the nonpolymorphic light chain component of the major histocompatibility class I complex. The physiological role of ␤ 2 m has been likened to a chaperone in that it is required for the heavy chain of the major histocompatibility class I complex to fold into a stable secreted entity (8, 9). ␤ 2 m is an all ␤-sheet protein, with a seven-stranded ␤-sandwich structure and a single disulfide bond linking Cys-25 and Cys-80 in the B and F strands (10) (Fig. 1). In healthy individuals, the serum concentration of circulating free ␤ 2 m is ϳ3 mg⅐liter Ϫ1 (11), and the protein is removed from the serum by renal catabolism (12). In individuals undergoing renal dialysis, the serum concentration of ␤ 2 m can rise to...

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“…The X-FEL was then focused down to a 0.1-m diameter spot (14) (which corresponds to 2.55 ϫ 10 6 photons͞Å 2 and is a value that is about at the state of the art). By using the spraying techniques from mass spectrometry (15,16), identical biomolecules can be selected and inserted one by one in random orientation into the X-FEL beam. With the simulated X-FEL exposures, we calculated 2D diffraction patterns from single biomolecules, each generated from a single pulse before the radiation damage manifests itself.…”

Section: Methodsmentioning

confidence: 99%

An approach to three-dimensional structures of biomolecules by using single-molecule diffraction images

Miao

Hodgson

Sayre

2001

Proc. Natl. Acad. Sci. U.S.A.

139

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We describe an approach to the high-resolution three-dimensional structural determination of macromolecules that utilizes ultrashort, intense x-ray pulses to record diffraction data in combination with direct phase retrieval by the oversampling technique. It is shown that a simulated molecular diffraction pattern at 2.5-Å resolution accumulated from multiple copies of single rubisco biomolecules, each generated by a femtosecondlevel x-ray free electron laser pulse, can be successfully phased and transformed into an accurate electron density map comparable to that obtained by more conventional methods. The phase problem is solved by using an iterative algorithm with a random phase set as an initial input. The convergence speed of the algorithm is reasonably fast, typically around a few hundred iterations. This approach and phasing method do not require any ab initio information about the molecule, do not require an extended ordered lattice array, and can tolerate high noise and some missing intensity data at the center of the diffraction pattern. With the prospects of the x-ray free electron lasers, this approach could provide a major new opportunity for the highresolution three-dimensional structure determination of single biomolecules.

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Mass spectrometry and viral analysis

Abstract: These results demonstrate the utility of electrospray for supramolecular complexes with molecular weights of over 40 million Da and offer conclusive evidence that native biomolecular structures can be conserved through the electrospray process.

Cited by 188 publications

References 18 publications

Nano ES GEMMA and PDMA, new tools for the analysis of nanobioparticles—Protein complexes, lipoparticles, and viruses

Nano ES GEMMA and PDMA, new tools for the analysis of nanobioparticles—Protein complexes, lipoparticles, and viruses

Co-populated Conformational Ensembles of β2-Microglobulin Uncovered Quantitatively by Electrospray Ionization Mass Spectrometry

An approach to three-dimensional structures of biomolecules by using single-molecule diffraction images

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