De novo sequencing of peptides on single resin beads by MALDI-FTICR tandem mass spectrometry
peer-reviewed · Journal of the American Society for Mass Spectrometry · 2010
| Date | 2010-02-01 |
| Type | peer-reviewed |
| Venue | Journal of the American Society for Mass Spectrometry |
| Publisher | American Chemical Society (ACS) |
| Contribution | adjacent |
| DOI | 10.1016/j.jasms.2009.10.004 |
| Citations (OpenAlex) | 20 |
| Venue 2-year citedness | 2.84 |
Abstract
An efficient approach in combinatorial chemistry is the synthesis of one-bead-one-compound peptide libraries. In contrast to synthesis and functional screening, which is performed in a largely automated manner, structure determination has been frequently laborious and time-consuming. Here we report an approach for de novo sequencing of peptides on single beads by matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance (MALDI-FTICR) tandem mass spectrometry, using a resin with a photolinker for solid-phase peptide synthesis. Upon sorting out single beads, an efficient sample preparation on the MALDI target was developed that enables fragmentation upon irradiation of the bead-matrix mixture with the ultraviolet (UV)-MALDI laser, with enhanced yield of sequence-specific fragment ions at increased laser energy. This approach is illustrated by sequence determinations of two peptides from a library with sequences varying in a single amino acid; the feasibility with tandem-MS procedures and fragment ion assignment was ascertained by sustained off-resonance irradiation/collision induced dissociation (SORI/CID) and infrared multiphoton dissociation (IRMPD) fragmentation.
Methods and tools
- Single-bead MALDI-FTICR de novo sequencing: Sequences peptides directly from single resin beads of one-bead-one-compound libraries by MALDI-FTICR tandem MS, using a photocleavable linker so the laser releases and fragments the peptide.