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

IF 1.3 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Journal of Biomolecular NMR Pub Date : 2024-03-21 DOI:10.1007/s10858-024-00440-z

Giorgia Toscano, Julian Holzinger, Benjamin Nagl, Georg Kontaxis, Hanspeter Kählig, Robert Konrat, Roman J. Lichtenecker

引用次数: 0

Abstract

We present an economic and straightforward method to introduce ¹³C-¹⁹F 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 [4-¹³C, 2,3,5,6-²H₄] 4-fluorophenylalanine from the commercially available isotope sources [2-¹³C] 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.

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用三重标记的 13C/19F/2H 同位素模式装饰苯丙氨酸侧链。

我们提出了一种经济而直接的方法，将 13C-19F 自旋系统引入苯丙氨酸的氚化芳香侧链，作为各种蛋白质核磁共振应用的报告物。该方法基于从市售同位素来源 [2-13C] 丙酮和氧化氘合成 [4-13C, 2,3,5,6-2H4] 4-氟苯丙氨酸。这种化合物很容易在含草甘膦的最小培养基中被标准大肠杆菌过表达代谢，从而在相应的目标蛋白质中产生高掺入率。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Biomolecular NMR 生物-光谱学

CiteScore

6.00

自引率

3.70%

发文量

审稿时长

6-12 weeks

期刊介绍： The Journal of Biomolecular NMR provides a forum for publishing research on technical developments and innovative applications of nuclear magnetic resonance spectroscopy for the study of structure and dynamic properties of biopolymers in solution, liquid crystals, solids and mixed environments, e.g., attached to membranes. This may include: Three-dimensional structure determination of biological macromolecules (polypeptides/proteins, DNA, RNA, oligosaccharides) by NMR. New NMR techniques for studies of biological macromolecules. Novel approaches to computer-aided automated analysis of multidimensional NMR spectra. Computational methods for the structural interpretation of NMR data, including structure refinement. Comparisons of structures determined by NMR with those obtained by other methods, e.g. by diffraction techniques with protein single crystals. New techniques of sample preparation for NMR experiments (biosynthetic and chemical methods for isotope labeling, preparation of nutrients for biosynthetic isotope labeling, etc.). An NMR characterization of the products must be included.