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Issue 17, 2011
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Evidence for α-helices in the gas phase: A case study using Melittin from honey bee venom

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Abstract

Gas phase methodologies are increasingly used to study the structure of proteins and peptides. A challenge to the mass spectrometrist is to preserve the structure of the system of interest intact and unaltered from solution into the gas phase. Small peptides are very flexible and can present a number of conformations in solution. In this work we examine Melittin a 26 amino acid peptide that forms the active component of honey bee venom. Melittin is haemolytic and has been shown to form an α-helical tetrameric structure by X-ray crystallography [M. Gribskov et al., The RCSB Protein Data Bank, 1990] and to be helical in high concentrations of methanol. Here we use ion mobility mass spectrometry, molecular dynamics and gas-phase HDX to probe its structure in the gas phase and specifically interrogate whether the helical form can be preserved. All low energy calculated structures possess some helicity. In our experiments we examine the peptide following nano-ESI from solutions with varying methanol content. Ion mobility gives collision cross sections (CCS) that compare well with values found from molecular modelling and from other reported structures, but with inconclusive results regarding the effect of solvent. There is only a slight increase in CCS with charge, showing minimal coloumbically driven unfolding. HDX supports preservation of some helical content into the gas phase and again shows little difference in the exchange rates of species sprayed from different solvents. The [M + 3H]3+ species has two exchanging populations both of which exhibit faster exchange rates than observed for the [M + 2H]2+ species. One interpretation for these results is that the time spent being analysed is sufficient for this peptide to form a helix in the ‘ultimate’ hydrophobic environment of a vacuum.

Graphical abstract: Evidence for α-helices in the gas phase: A case study using Melittin from honey bee venom

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Publication details

The article was received on 08 Apr 2011, accepted on 03 Jun 2011 and first published on 24 Jun 2011


Article type: Paper
DOI: 10.1039/C1AN15291B
Citation: Analyst, 2011,136, 3446-3452
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    Evidence for α-helices in the gas phase: A case study using Melittin from honey bee venom

    H. V. Florance, A. P. Stopford, J. M. Kalapothakis, B. J. McCullough, A. Bretherick and P. E. Barran, Analyst, 2011, 136, 3446
    DOI: 10.1039/C1AN15291B

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