Nonionic, Cleavable Surfactant for Top-Down Proteomics.
Kyle A BrownMorgan K GuggerZhen YuDavid MorenoSong JinYing GePublished in: Analytical chemistry (2023)
Nonionic surfactants are often used as general reagents for cell lysis enabling protein extraction, stabilization, and purification under nondenaturing conditions for downstream analysis in structural biology. However, the presence of surfactants in the sample matrix often has a deleterious effect on electrospray ionization (ESI)-mass spectrometry (MS) analysis of proteins and complexes. Here, we report a nonionic, cleavable surfactant, n-decyl-disulfide-β-D-maltoside (DSSM), for top-down proteomics. DSSM was designed to mimic the properties of one of the most common surfactants used in structural biology, n-dodecyl-β-D-maltoside (DDM), but contains a disulfide bond that allows for facile cleavage and surfactant removal before or during MS analysis. We have shown that DSSM is compatible with direct electrospray ionization (ESI)-MS analysis and reversed-phase liquid chromatography (RPLC)-MS analysis of proteins and protein complexes. We have demonstrated that DSSM can facilitate top-down proteomic characterization of membrane proteins such as a model ion channel protein and a G protein-coupled receptor as well as endogenous proteins from cell lysates for the determination of sequence variations and posttranslational modifications (PTMs). Conceivably, DSSM could serve as a general replacement for DDM in proteomic experiments and structural biology studies.
Keyphrases
- mass spectrometry
- liquid chromatography
- ms ms
- multiple sclerosis
- capillary electrophoresis
- gas chromatography
- single cell
- high performance liquid chromatography
- high resolution mass spectrometry
- high resolution
- protein protein
- amino acid
- label free
- solid phase extraction
- stem cells
- simultaneous determination
- bone marrow
- binding protein