Enzymatic stereospecific preparation of fluorescent S-adenosyl-L-methionine analogs.
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 K(d) of 0.38 microM for the S-2,6-diaminopurinylmethionine-SAM-III riboswitch complex.
Ottink
Otmar M
OM
Institute for Molecules and Materials, Department of Biophysical Chemistry, Radboud University Nijmegen, 6525 AJ Nijmegen, The Netherlands.
Nelissen
Frank H T
FH
Derks
Yvonne
Y
Wijmenga
Sybren S
SS
Heus
Hans A
HA
D029968:Escherichia coli Proteins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry*; D008716:Methionine Adenosyltransferase / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000031:analogs & derivatives* / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism
D029968:Escherichia coli Proteins; D005456:Fluorescent Dyes; D012436:S-Adenosylmethionine; C519385:MetK protein, E coli; D008716:Methionine Adenosyltransferase