Enzymatic stereospecific preparation of fluorescent S-adenosyl-l-methionine analogs |
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Authors: | Otmar M. Ottink |
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Affiliation: | Institute for Molecules and Materials, Department of Biophysical Chemistry, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands |
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Abstract: | S-Adenosyl-l-methionine (SAM) is the preferred cofactor for biological methyl group transfers to various substrates such as nucleic acids, proteins, and lipids. Here we present stereospecific (>95% of the desired enantiomer) and high-yield preparation of four fluorescent and biologically active SAM analogs and demonstrate their usefulness in binding studies. Using a fluorescence titration experiment, we obtained a Kd of 0.38 μM for the S-2,6-diaminopurinylmethionine-SAM-III riboswitch complex. |
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Keywords: | Enzyme catalysis Fluorescence spectroscopy Nucleotides RNA SAM Methylation |
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