| Info. | Vol.5 - No.3 (2011.09.20) |
|---|---|
| Title | Analysis of chemical/biochemical conversions on gold microparticles using MALDI-TOF MS |
| Authors | Jae Il Kim1, Sohyun Kim1,& Woon-Seok Yeo1 |
| Institutions | 1Department of Bioscience of Biotechnology, Konkuk University, Seoul 143-701, Korea Correspondence and requests for materials should be addressed to W.-S. Yeo ( wsyeo@konkuk.ac.kr) |
| Abstract | Chemical and biochemical analyses on solid supports are now in common because of the feasibility and simplicity of the process. For example, post-translational modifications of proteins such as phosphorylation, acetylation, and glycosylation have been assayed on several types of solid supports, mostly on 2-dimensional formats. Use of 3-dimensional solid supports, such as gold microparticles in this study, is beneficial over 2-dimensional biochips in terms of larger active surface areas and easier handling protocol. In this report, we present analyses of chemical conversions as well as enzymatic conversions on gold microparticles using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOFMS). For immobilization of enzyme substrates, gold microparticles, which presented organic functional groups, were subjected to a series of chemical reactions encompassing amide coupling and Michael addition. The resulting particles were then treated with enzymes. Direct analysis of the resulting products on microparticles by MALDI-TOF MS was performed without additional labeling steps, called SAMDI (self-assembled monolayers for MALDI). The mass spectra clearly showed chemical modifications and enzymatic conversions of peptide substrates on gold microparticles. We expect that our method can be used routinely in biological research, such as enzyme inhibitor assays, and development of substrate peptide of enzymes. We believe that the combination of gold microparticles with MALDI-TOF MS will provide an efficient analytical platform for use in various biochemical studies. |
| Keyword | Gold microparticles, Kinase, Mass spectrometry,Monolayers, Peptidase |
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