8PHL

Human carbonic anhydrase II containing 4-fluorophenylalanine

8PHL の概要

• エントリーDOI: 10.2210/pdb8phl/pdb
• 関連するPDBエントリー: 8B29 8P6U 8Q0C
• 分子名称: Carbonic anhydrase 2, MERCURIBENZOIC ACID, ZINC ION, ... (4 entities in total)
• 機能のキーワード: hcaii, carbonic anhydrase, 4-fluorophenylalanine, lyase
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 29673.78

構造登録者

Pham, L.B.T.,Costantino, A.,Barbieri, L.,Calderone, V.,Luchinat, E.,Banci, L. (登録日: 2023-06-20, 公開日: 2024-03-06)

主引用文献

Costantino, A.,Pham, L.B.T.,Barbieri, L.,Calderone, V.,Ben-Nissan, G.,Sharon, M.,Banci, L.,Luchinat, E.
Controlling the incorporation of fluorinated amino acids in human cells and its structural impact.
Protein Sci., 33:e4910-e4910, 2024

PubMed Abstract: Fluorinated aromatic amino acids (FAAs) are promising tools when studying protein structure and dynamics by NMR spectroscopy. The incorporation FAAs in mammalian expression systems has been introduced only recently. Here, we investigate the effects of FAAs incorporation in proteins expressed in human cells, focusing on the probability of incorporation and its consequences on the F NMR spectra. By combining F NMR, direct MS and x-ray crystallography, we demonstrate that the probability of FAA incorporation is only a function of the FAA concentration in the expression medium and is a pure stochastic phenomenon. In contrast with the MS data, the x-ray structures of carbonic anhydrase II reveal that while the 3D structure is not affected, certain positions lack fluorine, suggesting that crystallization selectively excludes protein molecules featuring subtle conformational modifications. This study offers a predictive model of the FAA incorporation efficiency and provides a framework for controlling protein fluorination in mammalian expression systems.

PubMed: 38358125
DOI: 10.1002/pro.4910

実験手法

X-RAY DIFFRACTION (1.3 Å)