Structure of melittin bound to phospholipid micelles studied using hydrogen-deuterium exchange and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.
The structure of melittin bound to dodecylphosphocholine (DPC) micelles was investigated using hydrogen-deuterium (H/D) exchange in conjunction with collision induced dissociation (CID) in an rf-only hexapole ion guide with electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS). The deuterium incorporation into backbone amide hydrogens of melittin with or without DPC micelles was analyzed at different time points examining the mass of each fragment ion produced by hexapole CID. When melittin existed alone in aqueous solution, more than 80% of amide hydrogens was exchanged within 10 s, and the deuterium content in each fragment ion showed high values throughout the experiments. When melittin was bound to DPC micelles, the percentage of deuterium incorporation into the fragment decreased remarkably at any time point. It increased little by little as the exchange period prolonged, indicating that some stable structure was formed by the interaction with DPC. The results obtained here were consistent with the previous studies on the helical structure of melittin carried out by NMR and CD analyses. The strategy using H/D exchange and MS analysis might be useful for studying structural changes of peptides and proteins caused by phospholipid micelles. It could also be applied to membrane-bound proteins to characterize their structure.
Akashi
S
S
Division of Biomolecular Characterization, RIKEN (The Institute of Physical and Chemical Research), Saitama, Japan. akashi@tsurumi.yokohama-cu.ac.jp
Takio
K
K
D000818:Animals; D001516:Bees / Q000737:chemistry; D017357:Cyclotrons; D003903:Deuterium / Q000737:chemistry; D005583:Fourier Analysis; D006859:Hydrogen / Q000737:chemistry*; D008555:Melitten / Q000737:chemistry*; D008823:Micelles; D010743:Phospholipids / Q000737:chemistry*; D011487:Protein Conformation; D021241:Spectrometry, Mass, Electrospray Ionization
D008823:Micelles; D010743:Phospholipids; D008555:Melitten; D006859:Hydrogen; D003903:Deuterium