Hydrogen/deuterium exchange of hydrophobic peptides in model membranes by electrospray ionization mass spectrometry

Hansen, Raino K.; Broadhurst, R. William; Skelton, Paul; Arkin, Isaiah T.

doi:10.1016/s1044-0305(02)00702-x

Cited by 28 publications

(20 citation statements)

References 54 publications

(65 reference statements)

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“…4°C (Figure 3), the rapid 37°C thaw was chosen for further work. This thaw is equivalent to that reported in a recent paper on plasmid vectors (Armstrong and Anchordoquy 2004) and liposomes (Hansen et al 2002).…”

Section: Resultssupporting

confidence: 85%

Mechanisms of Inactivation of HSV-2 during Storage in Frozen and Lyophilized Forms

Hansen

Zhai

Skepper

et al. 2008

Biotechnol Progress

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The structural integrity of herpes simplex virus 2 (HSV-2) during freezing, thawing, and lyophilization has been studied using scanning and transmission electron microscopy. Viral particles should be thawed quickly from -80 to 37 degrees C to avoid artifacts of thawing. To avoid freezing damage, the virus should be rapidly frozen (>20 K s(-1)) rather than slowly frozen as occurs on the shelf of a lyophilizer (<1 K s(-1)). Fast freezing and thawing allows six cycles of freeze thaw with no loss of viral titer TCID50. Viral particles were characterized using immunogold labeling methods. Freshly thawed virus had 19 +/- 4 polyclonal immunogold particles virus(-1); virus stored at -80 degrees C for at least 4 months had 17 +/- 3 particles virus(-1); virus stored for 1 week at 4 degrees C had 8 +/- 4 particles virus(-1). By bulk lyophilization the number of particles was 4 +/- 4, but by fast freezing and lyophilization the number of gold particles improved to 12 +/- 5. The loss of viral membrane was directly observed, and the in vitro loss was demonstrated to occur through three possible pathways, including (i) simultaneous release of tegument and membrane, (ii) sequential release of membrane and then tegument, and (iii) release like by in vivo infection. The capsids were not further degraded as indicated by the lack of free DNA, which was only released by boiling the viral samples with 1% SDS, followed by a dilution to 0.001% w/v SDS for the real-time PCR reaction.

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Section: Resultssupporting

confidence: 85%

Mechanisms of Inactivation of HSV-2 during Storage in Frozen and Lyophilized Forms

Hansen

Zhai

Skepper

et al. 2008

Biotechnol Progress

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“…Such receptors could be examined with regard to the extent of amide backbone NH to N 2 H exchange, both in the presence and absence of ligand. 53 Reduced exchange indicates structural tightening. 1,15 However, it is already clear from the systems that have been so far studied that differences in the biological effects of different ligands at receptors are related to fundamental thermodynamic considerations.…”

Section: Resultsmentioning

confidence: 99%

Order Changes within Receptor Systems upon Ligand Binding: Receptor Tightening/Oligomerisation and the Interpretation of Binding Parameters

Williams

Sandercock

Stephens

2004

Journal of Molecular Biology

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Recent hydrogen-deuterium exchange experiments have highlighted tightening and loosening of protein structures upon ligand binding, with changes in bonding (DeltaH) and order (DeltaS) which contribute to the overall thermodynamics of ligand binding. Tightening and loosening show that ligand binding respectively stabilises or destabilises the internal structure of the protein, i.e. it shows positive or negative cooperativity between ligand binding and the receptor structure. In the case of membrane-bound receptors, such as G protein-coupled receptors (GPCRs) and ligand gated ion channel receptors (LGICRs), most binding studies have focussed on association/dissociation constants. Where these have been broken down into enthalpic and entropic contributions, the phenomenon of "thermodynamic discrimination" between antagonists and agonists has often been noted; e.g. for a receptor where agonist binding is predominantly enthalpy driven, antagonist binding is predominantly entropy driven and vice versa. These data have not previously been considered in terms of the tightening, or loosening, of receptor structures that respectively occurs upon positively, or negatively, cooperative binding of ligand. Nor have they been considered in light of the homo- and hetero-oligomerisation of GPCRs and the possibility of ligand-induced changes in oligomerisation. Here, we argue that analysis of the DeltaH and DeltaS of ligand binding may give useful information on ligand-induced changes in membrane-bound receptor oligomers, relevant to the differing effects of agonists and antagonists.

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“…Peptide concentrations are estimated as approximately 2 μM. Samples were analyzed by direct‐infusion electrospray after mixing for 2 min in the deuterated solution 6…”

Section: Methodsmentioning

confidence: 99%

Investigation of apparent mass deviations in electrospray ionization tandem mass spectrometry of a benzophenone‐labeled peptide

Leite

Dougherty

Lester

et al. 2003

Rapid Comm Mass Spectrometry

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In a previous study utilizing benzophenone-based topological probes to study conformationally dependent changes in mouse muscle nicotinic acetylcholine receptor (nAChR) topology, electrospray ionization tandem mass spectrometric (ESI-MS/MS) analysis led to a consistent -2.0 Da mass deviation from expected values. In the present study a synthetic peptide, corresponding to nAChR alpha1 subunit residues 130-139, was photolabeled. MS/MS analysis of this peptide using an ion trap confirmed the previously observed mass deviation, associated only with fragment ions that contain the incorporated benzophenone moiety. Analysis of peak profiles for the photolabeled ions does not indicate the typical 'peak fronting' that produces a mass shift when labile ions are prematurely ejected from the ion trap. Rather, hydrogen/deuterium (H/D) exchange experiments support the hypothesis that a chemical rearrangement involving phenyl migration and ketone formation has formed an unexpected oxidized peptide, with molecular mass 2 Da less than that expected, that is isolated for collision-induced dissociation in the ion trap together with the predicted precursor due to the broad ion isolation window specified.

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Hydrogen/deuterium exchange of hydrophobic peptides in model membranes by electrospray ionization mass spectrometry

Cited by 28 publications

References 54 publications

Mechanisms of Inactivation of HSV-2 during Storage in Frozen and Lyophilized Forms

Mechanisms of Inactivation of HSV-2 during Storage in Frozen and Lyophilized Forms

Order Changes within Receptor Systems upon Ligand Binding: Receptor Tightening/Oligomerisation and the Interpretation of Binding Parameters

Investigation of apparent mass deviations in electrospray ionization tandem mass spectrometry of a benzophenone‐labeled peptide

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