| Combination of Mixed Self-Assembled Monolayer and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry, a Simple Tip-Based Screening Method for Proteomics |
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1 Protein Research Group, RIKEN Genomic Sciences Center, Yokohama, JAPAN 2 Department of Chemistry, School of Science, The University of Tokyo, TOKYO, JAPAN 3 Division of Supramolecular Biology, Yokohama City University, Yokohama, JAPAN |
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| A new screening method to detect protein-ligand interactions has been developed through a combination of mixed self-assembled monolayer (SAM) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The mixed SAM surface was prepared on flat gold plate from a mixture of 20-mercapto-3,6,9-trioxaicosanol and a ligand-bound derivative of 29-mercapto-3,6,9,12,15,18-hexaoxanonacosanol. This surface is very suitable for observing the molecular interaction accurately, because there is very low levels of steric hindrance between immobilized ligands and the non-charged hydroxyl groups of polyethylene glycol which resists from the non-specific adsorption of biomolecules. Because the surface coated with the mixed SAM is washable with water, it can be used directly as a probe for MALDI-MS, a technique commonly used for the annotation of protein function. As an example we report here the selective detection of the molecular interaction between a Src homology 3 (SH3) domain of human p85α phosphatidylinositol 3-kinase (PI3K) and its ligand peptide (H-RKLPPRPAF-OH) attached covalently to mixed SAM using MALDI-MS. | ||
| Key words: Self-assembled monolayer, Functionalized thiol, MALDI-MS, Affinity capillary electrophoresis, PI3K-SH3 domain | ||
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