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1
The crystallization of apo-form UMP kinase from Xanthomonas campestris is significantly improved in a strong magnetic field.来自野油菜黄单胞菌的脱辅基形式的UMP激酶在强磁场中的结晶得到显著改善。
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 May 1;63(Pt 5):438-42. doi: 10.1107/S1744309107018787. Epub 2007 Apr 20.
2
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3
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Biochim Biophys Acta. 2005 Mar 14;1747(2):271-5. doi: 10.1016/j.bbapap.2004.11.010. Epub 2004 Dec 9.
4
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The crystal structure of Pyrococcus furiosus UMP kinase provides insight into catalysis and regulation in microbial pyrimidine nucleotide biosynthesis.嗜热栖热菌尿苷一磷酸激酶的晶体结构为深入了解微生物嘧啶核苷酸生物合成中的催化作用和调节机制提供了线索。
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Structural and enzymatic investigation of the Sulfolobus solfataricus uridylate kinase shows competitive UTP inhibition and the lack of GTP stimulation.嗜热栖热菌尿苷酸激酶的结构与酶学研究表明存在竞争性UTP抑制且缺乏GTP刺激。
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8
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A putative polyketide-synthesis protein XC5357 from Xanthomonas campestris: heterologous expression, crystallization and preliminary X-ray analysis.来自野油菜黄单胞菌的一种假定聚酮化合物合成蛋白XC5357：异源表达、结晶及初步X射线分析。
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Crystallization and preliminary X-ray analysis of XC1015, a histidine triad-like protein from Xanthomonas campestris.野油菜黄单胞菌中一种组氨酸三联体样蛋白XC1015的结晶及X射线初步分析
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本文引用的文献

1
[10] Dynamic light scattering in evaluating crystallizability of macromolecules.[10] 动态光散射在评估大分子结晶性中的应用
Methods Enzymol. 1997;276:157-166. doi: 10.1016/S0076-6879(97)76056-7.
2
Processing of X-ray diffraction data collected in oscillation mode.振荡模式下收集的X射线衍射数据的处理。
Methods Enzymol. 1997;276:307-26. doi: 10.1016/S0076-6879(97)76066-X.
3
Assessment of a preliminary solubility screen to improve crystallization trials: uncoupling crystal condition searches.评估初步溶解度筛选以改进结晶试验：解开晶体条件搜索。
Acta Crystallogr D Biol Crystallogr. 2006 Jul;62(Pt 7):833-42. doi: 10.1107/S0907444906018385. Epub 2006 Jun 20.
4
Crystallization Optimum Solubility Screening: using crystallization results to identify the optimal buffer for protein crystal formation.结晶最佳溶解度筛选：利用结晶结果确定蛋白质晶体形成的最佳缓冲液。
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Dec 1;61(Pt 12):1035-8. doi: 10.1107/S1744309105035244. Epub 2005 Nov 5.
5
Cloning, purification, crystallization and preliminary X-ray crystallographic analysis of XC847, a 3'-5' oligoribonuclease from Xanthomonas campestris.野油菜黄单胞菌3'-5'寡核糖核酸酶XC847的克隆、纯化、结晶及初步X射线晶体学分析
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Oct 1;61(Pt 10):902-5. doi: 10.1107/S1744309105027132. Epub 2005 Sep 30.
6
The crystal structure of Pyrococcus furiosus UMP kinase provides insight into catalysis and regulation in microbial pyrimidine nucleotide biosynthesis.嗜热栖热菌尿苷一磷酸激酶的晶体结构为深入了解微生物嘧啶核苷酸生物合成中的催化作用和调节机制提供了线索。
J Mol Biol. 2005 Sep 16;352(2):438-54. doi: 10.1016/j.jmb.2005.07.045.
7
Effects of Tween 20 and Tween 80 on the stability of Albutropin during agitation.吐温20和吐温80对沙丁胺醇在搅拌过程中稳定性的影响。
J Pharm Sci. 2005 Jun;94(6):1368-81. doi: 10.1002/jps.20365.
8
Structure of Escherichia coli UMP kinase differs from that of other nucleoside monophosphate kinases and sheds new light on enzyme regulation.大肠杆菌尿苷一磷酸激酶的结构不同于其他核苷单磷酸激酶，为酶的调控提供了新线索。
J Biol Chem. 2005 Jul 8;280(27):25533-40. doi: 10.1074/jbc.M501849200. Epub 2005 Apr 27.
9
Structural consequences of hen egg-white lysozyme orthorhombic crystal growth in a high magnetic field: validation of X-ray diffraction intensity, conformational energy searching and quantitative analysis of B factors and mosaicity.高磁场中蛋清溶菌酶正交晶体生长的结构后果：X射线衍射强度的验证、构象能量搜索以及B因子和镶嵌性的定量分析
Acta Crystallogr D Biol Crystallogr. 2005 Mar;61(Pt 3):207-17. doi: 10.1107/S0907444904030926. Epub 2005 Feb 24.
10
Optimum solubility (OS) screening: an efficient method to optimize buffer conditions for homogeneity and crystallization of proteins.最佳溶解度（OS）筛选：一种优化蛋白质均一性和结晶缓冲条件的有效方法。
Acta Crystallogr D Biol Crystallogr. 2004 Sep;60(Pt 9):1670-3. doi: 10.1107/S0907444904010972. Epub 2004 Aug 26.

来自野油菜黄单胞菌的脱辅基形式的UMP激酶在强磁场中的结晶得到显著改善。

The crystallization of apo-form UMP kinase from Xanthomonas campestris is significantly improved in a strong magnetic field.

作者信息

Tu Jhe-Le, Chin Ko-Hsin, Wang Andrew H-J, Chou Shan-Ho

机构信息

Institute of Biochemistry, National Chung-Hsing University, Taichung 40227, Taiwan.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2007 May 1;63(Pt 5):438-42. doi: 10.1107/S1744309107018787. Epub 2007 Apr 20.

DOI:10.1107/S1744309107018787
PMID:17565191
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2335002/
Abstract

Bacterial UMP kinases (UMPKs) are crucial enzymes that are responsible for microbial UTP biosynthesis. Interestingly, eukaryotic and prokaryotic cells use different enzymes for UMP-phosphorylation reactions. Prokaryotic UMPKs are thus believed to be potential targets for antimicrobial drug development. Here, the cloning, expression and crystallization of SeMet-substituted XC1936, a bacterial UMPK from Xanthomonas campestris pathovar campestris, are reported. The crystallization of the apo-form UMPK was found to be significantly improved in a strong magnetic field; the crystals diffracted to a resolution of 2.35 A, a dramatic improvement over the original value of 3.6 A. Preliminary structural analyses of apo-form XC1936 using crystals grown in a strong magnetic field clearly reveal well defined loop regions involved in substrate-analogue binding that were previously not visible. Crystallization in a strong magnetic field thus was found to be indispensable in determining the flexible region of the XC1936 UMPK structure.

摘要

细菌尿苷一磷酸激酶（UMPK）是负责微生物三磷酸尿苷（UTP）生物合成的关键酶。有趣的是，真核细胞和原核细胞在UMP磷酸化反应中使用不同的酶。因此，原核UMPK被认为是抗菌药物开发的潜在靶点。本文报道了来自野油菜黄单胞菌野油菜致病变种的细菌UMPK——硒代甲硫氨酸取代的XC1936的克隆、表达和结晶。发现无配体形式的UMPK在强磁场中结晶得到显著改善；晶体衍射分辨率达到2.35 Å，相较于原来的3.6 Å有了显著提高。使用在强磁场中生长的晶体对无配体形式的XC1936进行的初步结构分析清楚地揭示了以前不可见的参与底物类似物结合的明确环区域。因此，发现在强磁场中结晶对于确定XC1936 UMPK结构的柔性区域是必不可少的。