Profiling Substrate Promiscuity of Wild-Type Sugar Kinases for Multi-fluorinated Monosaccharides.
Profiling Substrate Promiscuity of Wild-Type Sugar Kinases for Multi-fluorinated Monosaccharides.
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DOI：
10.26434/chemrxiv.11950647.v1
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发表时间：
2020-03
影响因子：
8.6
通讯作者：
T. Keenan;F. Parmeggiani;J. Malassis;C. Fontenelle;J. Vendeville;W. Offen;P. Both;Kun Huang;A. Marchesi;A. Heyam;C. Young;S. Charnock;G. Davies;B. Linclau;S. Flitsch;M. Fascione
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作者：
T. Keenan;F. Parmeggiani;J. Malassis;C. Fontenelle;J. Vendeville;W. Offen;P. Both;Kun Huang;A. Marchesi;A. Heyam;C. Young;S. Charnock;G. Davies;B. Linclau;S. Flitsch;M. Fascione
氟化糖-1-磷酸作为修饰寡糖的化学和生物催化合成的中间体以及化学生物学探针具有新兴的重要性。在这里，我们提出了一个系统的研究，对一个面板的氟化单糖的异头糖激酶（半乳糖和N-乙酰己糖胺激酶）的广泛的活性，导致第一个例子的多氟化底物接受这类酶。我们已经发现了四个新的N-乙酰己糖胺激酶具有不同的底物范围，从而扩大了这一亚类激酶的同系物的数量。最后，我们已经解决了与2-脱氧-2-氟半乳糖复合的半乳糖激酶的晶体结构，从而深入了解活性位点的变化，这可能是酶对某些底物类似物的特异性的原因。
Fluorinated sugar-1-phosphates are of emerging importance as intermediates in the chemical and biocatalytic synthesis of modified oligosaccharides, as well as probes for chemical biology. Here we present a systematic study of the activity of a wide range of anomeric sugar kinases (galacto- and N-acetylhexosamine kinases) against a panel of fluorinated monosaccharides, leading to the first examples of polyfluorinated substrates accepted by this class of enzymes. We have discovered four new N-acetylhexosamine kinases with a different substrate scope, thus expanding the number of homologs available in this subclass of kinases. Lastly, we have solved the crystal structure of a galactokinase in complex with 2-deoxy-2-fluorogalactose, giving insight into changes in the active site that may account for the specificity of the enzyme toward certain substrate analogs.