title: Separation of peptides from detergents using ion mobility spectrometry.
abstract: Mass spectrometry (MS) has dramatically evolved in the last two decades and has been the driving force of the spectacular expansion of proteomics during this period. However, the very poor compatibility of MS with detergents is still a technical obstacle in some studies, in particular on membrane proteins. Indeed, the high hydrophobicity of membrane proteins necessitates the use of detergents for their extraction and solubilization. Here, we address the analytical potential of high-field asymmetric waveform ion mobility spectrometry (FAIMS) for separating peptides from detergents. The study was focused on peptides from the human integral membrane protein CD9. A tryptic peptide was mixed with the non-ionic detergents Triton X-100 or beta-D-dodecyl maltoside (DDM) as well as with the ionic detergents sodium dodecyl sulfate (SDS) or sodium deoxycholate (SDC). Although electrospray ionization (ESI) alone led to a total suppression of the peptide ion signal on mass spectra with only detection of the detergents, use of FAIMS allowed separation and clear identification of the peptide with any of the detergents studied. The detection and identification of the target compound in the presence of an excess of detergents are then feasible. FAIMS should prove especially useful in the structural and proteomic analysis of membrane proteins.
authors: Inserm, U785, F-94800 Villejuif, France.
authors: Aïcha
authors: A
authors: Bagag
authors: Alexandre
authors: A
authors: Giuliani
authors: Francis
authors: F
authors: Canon
authors: Matthieu
authors: M
authors: Réfrégiers
authors: François
authors: F
authors: Le Naour
mesh_terms: D003840:Deoxycholic Acid / Q000737:chemistry; D003902:Detergents / Q000737:chemistry*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D017830:Octoxynol / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D060245:Tetraspanin 29 / Q000737:chemistry / Q000302:isolation & purification; D014357:Trypsin / Q000737:chemistry
chemical_list: C554869:CD9 protein, human; D003902:Detergents; D010446:Peptide Fragments; D060245:Tetraspanin 29; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol; D014357:Trypsin