Synthesis and incorporation of [6,7]-selenatryptophan into dihydrofolate reductase

被引:15
作者
Boles, JO [1 ]
Henderson, J
Hatch, D
Silks, LA
机构
[1] Tennessee Technol Univ, Dept Chem, Cookeville, TN 38505 USA
[2] Los Alamos Natl Lab, Biosci Div, Los Alamos, NM 87545 USA
关键词
beta-seleno[3,2-b]pyrrolyl-L-alanine; 4,5]selenatryptophan; beta-selenolo[2,3-b]pyrrolyl-L-alanine; 6,7]selenatryptophan; selenolo[2,3-b]pyrrole; selenomethionine; telluromethionine; 4,5]SeTrp; 6,7]SeTrp;
D O I
10.1016/S0006-291X(02)02438-5
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Until recently, the only selenium containing amino acid which could be used to completely substitute for a-wild type amino acid was selenomethionine (SeMet). In the last decade the preparation of SeMet containing proteins has proved to be valuable tools in the determination of three-dimensional structure by multiwavelength anomalous diffraction (MAD) techniques. The potential utility of a selenium containing tryptophan analog, beta-seleno[3,2-b]pyrrolyl-L-alanine ([4,5]selenatryptophan), has recently been demonstrated in the literature. This finding shows promise for the bioincorporation of its positional isomer, beta-selenolo[2,3-b]pyrrolyl-L-alanine ([6,7]selenatryptophan), thereby adding to the essential arsenal of selenium-containing amino acids for use in the characterization of proteins. The synthesis of [6,7]selenatryptophan by enzymatic biotransformation with tryptophan synthase from selenolo [2,3-b]pyrrole was carried out as well as its characterization by NMR spectroscopy and thin layer chromatography. Selenatryptophyl dihydrofolate reductase ([6,7]SeTrp-DHFR) was then synthesized in vivo, purified, and found to exhibit no perturbations to enzymatic activity. (C) 2002 Elsevier Science (USA). All rights reserved.
引用
收藏
页码:257 / 261
页数:5
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