Decorating phenylalanine side-chains with triple labeled 13C/19F/2H isotope patterns

被引:3
|
作者
Toscano, Giorgia [1 ,3 ]
Holzinger, Julian [2 ]
Nagl, Benjamin [4 ]
Kontaxis, Georg [5 ]
Kaehlig, Hanspeter [4 ]
Konrat, Robert [2 ]
Lichtenecker, Roman J. [1 ,6 ]
机构
[1] Univ Vienna, Inst Organ Chem, Christian Doppler Lab High Content Struct Biol & B, Wahringer Str 38, A-1090 Vienna, Austria
[2] Univ Vienna, Dept Struct & Computat Biol, Christian Doppler Lab High Content Struct Biol & B, Max Perutz Labs, Campus Vienna Bioctr 5, A-1030 Vienna, Austria
[3] Univ Vienna, Vienna Doctoral Sch Chem DoSChem, Wahringer Str 42, A-1090 Vienna, Austria
[4] Univ Vienna, Inst Organ Chem, Wahringer Str 38, A-1090 Vienna, Austria
[5] Univ Vienna, Max Perutz Labs, Dept Struct & Computat Biol, Campus Vienna Bioctr 5, A-1030 Vienna, Austria
[6] MAG LAB, Karl Farkas Gasse 22, A-1030 Vienna, Austria
关键词
Fluorine NMR; Fluorophenylalanine; Isotope labeling; F-TROSY; Protein overexpression; AMINO-ACIDS; F-19; NMR; RELAXATION; PROTEINS; TROSY;
D O I
10.1007/s10858-024-00440-z
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
We present an economic and straightforward method to introduce C-13-F-19 spin systems into the deuterated aromatic side chains of phenylalanine as reporters for various protein NMR applications. The method is based on the synthesis of [4-C-13, 2,3,5,6-H-2(4)] 4-fluorophenylalanine from the commercially available isotope sources [2-C-13] acetone and deuterium oxide. This compound is readily metabolized by standard Escherichia coli overexpression in a glyphosate-containing minimal medium, which results in high incorporation rates in the corresponding target proteins.
引用
收藏
页码:139 / 147
页数:9
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