Observation of a small oligonucleotide duplex by electrospray ionization mass spectrometry

Light-Wahl, Karen J.; Springer, David L.; Winger, Brian E.; Edmonds, Charles G.; Camp, David G.; Thrall, Brian D.; Smith, Richard D.

doi:10.1021/ja00055a070

Cited by 170 publications

(99 citation statements)

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“…It is not disrupted by guanidine hydrochloride, pH extremes, SDS, reducing agents or heat and it is resistant to many proteases [14]. Electrospray ionization mass spectrometry (ESMS) [15] has previously been used to characterise non-covalent receptor-ligand complexes [16], oligonucleotide associations [17,18], non-covalent protein-peptide complexes [19] and more recently to study the tetramers formed by avidin [20], streptavidin [21], hemoglobin [20] as well as yeast alcohol dehydrogenase and rabbit muscle pyruvate kinase [22]. However, the use of ESMS is usually limited to mass measurements below 150 kDa due to the difficulty of resolving multiply charged ions at higher mass.…”

Section: Introductionmentioning

confidence: 99%

Characterisation of the heptameric pore‐forming complex of the Aeromonas toxin aerolysin using MALDI‐TOF mass spectrometry

Moniatte¹,

Goot

Buckley

et al. 1996

FEBS Letters

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Aerolysin, a virulence factor secreted by Aeromonas hydrophila, is representative of a group of ~-sheet toxins that must form stable homooligomers in order to be able to insert into biological membranes and generate channels. Electron microscopy and image analysis of two-dimensional membrane crystals had previously revealed a structure with 7-fold symmetry suggesting that aerolysin forms heptameric oligomers [Wilmsen et al. (1992) EMBO J. 11, 2457-24631. However, this unusual molecularity of the channel remained to be confirmed by an independent method since low-resolution electron crystallography had led to artefactual data for other pore-forming toxins. In this study, matrix-assisted laser desorption/ionization time-offlight mass spectrometry (MALDI-TOF-MS) was used to measure the mass of the aerolysin oligomer preparation. A mass of 333 850 Da was measured, fitting very well with a beptameric complex (expected mass: 332 300 Da). These results confirm the earlier evidence that the aerolysin oligomer is a heptamer and also show that MALDI-TOF mass spectrometry could be a valuable tool to study non-covalent association of proteins.

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Section: Introductionmentioning

confidence: 99%

Characterisation of the heptameric pore‐forming complex of the Aeromonas toxin aerolysin using MALDI‐TOF mass spectrometry

Moniatte¹,

Goot

Buckley

et al. 1996

FEBS Letters

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“…ESI-MS, in particular, is increasingly being used to examine noncovalent complexes between protein subunits or proteins and their ligands. ESI-MS conditions can be mild enough that proteins retain their native conformations and enzyme-substrate complexes or subunit interactions can be observed (Ganem et al, 1991a(Ganem et al, , 1991bKatta & Chait, 1991b;Baca & Kent, 1992;Ganguly et al, 1992;Drummond et al, 1993;Goodlett et al, 1993;Light-Wahl et al, 1993aLoo et al, 1993aLoo et al, , 1993bOgorzalek Loo et al, 1993). A number of investigators have taken advantage of the dramatic increase in charge state observed when some proteins are denatured to probe protein structure (Katta & Chait, 1991aLeBlanc et al, 1991;Loo et al, 1991Loo et al, , 1993b Mirzaet al, 1993).…”

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confidence: 99%

Surfactant effects on protein structure examined by electrospray ionization mass spectrometry

1994

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Electrospray ionization mass spectrometry (ESI-MS) has proven to be a useful tool for examining noncovalent complexes between proteins and a variety of ligands. It has also been used to distinguish between denatured and refolded forms of proteins. Surfactants are frequently employed to enhance solubilization or to modify the tertiary or quaternary structure of proteins, but are usually considered incompatible with mass spectrometry. A broad range of ionic, nonionic, and zwitterionic surfactants was examined to characterize their effects on ESI-MS and on protein structure under ESI-MS conditions. Solution conditions studied include 4% acetic acid/50% acetonitrile/46% H 2 0 and 100% aqueous. Of the surfactants examined, the nonionic saccharides, such as n-dodecyl-P-D-glucopyranoside, at 0.1 Vo to 0.01% (w/v) concentrations, performed best, with limited interference from chemical background and adduct formation. Under the experimental conditions used, ESI-MS performance in the presence of surfactants was found to be unrelated to critical micelle concentration. It is demonstrated that surfactants can affect both the tertiary and quaternary structures of proteins under conditions used for ESI-MS. However, several of the surfactants caused significant shifts in the charge-state distributions, which appeared to be independent of conformational effects. These observations suggest that surfactants, used in conjunction with ESI-MS, can be useful for protein structure studies, if care is used in the interpretation of the results.Keywords: conformation; denaturation; detergent; electrospray; mass spectrometry; surfactant Surfactants play a significant role in protein chemistry with primary applications ranging from solubilization and stabilization of proteins to disaggregation of protein complexes and denaturation (Neugebauer, 1990(Neugebauer, , 1992Bollag & Edelstein, 1991). Some surfactants can disrupt protein higher order structure, whereas others are used as aids in protein refolding. They are employed in both gel and capillary electrophoresis and enhance protein and peptide recoveries from synthetic membranes employed in electroblotting and in electroelution of proteins from gels (Fernandez et al., 1994). Membrane-bound proteins, in particular, require surfactant treatment for solubilization. Abbreviotiom: CMC, critical micelle concentration; ESI, electrospray ionization; ESI-MS, electrospray ionization mass spectrometry; MALDI, matrix-assisted laser desorption/ionization; NP40, Nonidet P a , CHAPS, 3-(3-cholamidopropyl)dimethylammonio-I-propane sulfonate; CTAB, cetyl trimethylammonium bromide; LDAO, laurel dimethylamine oxide; n-dodecyl glucoside, n-dodecyl-0-D-glucopyranoside; n-hexylglucoside, n-hexyl-0-D-glucopyranoside; octyl glucoside, octyl-fl-D-glucopyranoside; octyl thioglucoside, I-S-octyl-fi-~-thioglucopyranoside.Matrix-assisted laser desorption/ionization or electrospray ionization have demonstrated molecular weight determinations for proteins greater than 100 kDa (Fenn et al., 1989;Smith et al., 19...

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“…The advent of soft ionization techniques and the advances of analyzers design have made this possibility a reality. Indeed, relatively short duplexes formed by complementary deoxyoligonucleotides were among the first noncovalent complexes detected by ESI without unwelcome dissociation [95][96][97]. Although this desirable outcome can be obtained also by MALDI with a judicious selection of matrix, additives, and other experimental conditions [98 -101], ESI remains the ionization technique of choice for the detection of nucleic acid noncovalent assemblies (reviewed in references [102][103][104][105]).…”

Section: Elucidating Structure-function Relationshipsmentioning

confidence: 99%

A eole for the MS analysis of nucleic acids in the post-genomics age

Fabris

2010

J. Am. Soc. Mass Spectrom.

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The advances of mass spectrometry in the analysis of nucleic acids have tracked very closely the exciting developments of instrumentation and ancillary technologies, which have taken place over the years. However, their diffusion in the broader life sciences community has been and will be linked to the ever evolving focus of biomedical research and its changing demands. Before the completion of the Human Genome Project, great emphasis was placed on sequencing technologies that could help accomplish this project of exceptional scale. After the publication of the human genome, the emphasis switched toward techniques dedicated to the exploration of sequences not coding for actual protein products, which amount to the vast majority of transcribed elements. The broad range of capabilities offered by mass spectrometry is rapidly advancing this platform to the forefront of the technologies employed for the structure-function investigation of these noncoding elements. Increasing focus on the characterization of functional assemblies and their specific interactions has prompted a re-evaluation of what has been traditionally construed as nucleic acid analysis by mass spectrometry. Inspired by the accelerating expansion of the broader field of nucleic acid research, new applications to fundamental biological studies and drug discovery will help redefine the evolving role of MS-analysis of nucleic acids in the post-genomics age. (J Am Soc Mass Spectrom 2010, 21, 1-13)

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Observation of a small oligonucleotide duplex by electrospray ionization mass spectrometry

Cited by 170 publications

References 0 publications

Characterisation of the heptameric pore‐forming complex of the Aeromonas toxin aerolysin using MALDI‐TOF mass spectrometry

Characterisation of the heptameric pore‐forming complex of the Aeromonas toxin aerolysin using MALDI‐TOF mass spectrometry

Surfactant effects on protein structure examined by electrospray ionization mass spectrometry

A eole for the MS analysis of nucleic acids in the post-genomics age

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