Solution structures of chemoenzymatically synthesized heparin and its precursors.
Solution structures of chemoenzymatically synthesized heparin and its precursors.
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DOI：
10.1021/ja8026345
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发表时间：
2008-10-01
影响因子：
15
通讯作者：
Linhardtt, Robert J.
中科院分区：
文献类型：
--
作者：
Zhang, Zhenqing;McCallum, Scott A.;Xie, Jin;Nieto, Lidia;Corzana, Francisco;Jimenez-Barbero, Jesus;Chen, Miao;Liu, Jian;Linhardtt, Robert J.
我们报道了首次从大肠杆菌K5中制备的均匀标记的[13C，15N] n -乙酰肝素（- glca (1,4)GlcNAc-）化学酶合成稳定同位素富集的肝素。糖胺聚糖（GAG）前体和肝素由n -乙酰型肝磷脂经过以下步骤形成：化学n -去乙酰化和n -磺化生成n -磺化型肝磷脂（- glca (1,4)GlcNS-）；酶催化的c5 -外映体化和2- o -磺化导致过硫酸化肝素（- idoa2s (1,4)GlcNS-）；酶促6- o -磺化导致肝素主链（- idoa2s (1,4)GlcNS6S-）；选择性酶促3- o磺化生成抗凝血肝素，含有GlcNS6S3S残基。采用异核、多维核磁共振波谱分析了[13C，15N] n -乙酰肝磷脂、前体和肝素的化学组成和溶液结构。发现同位素富集提供了分辨率高的13C光谱，具有这些生物分子构象研究所需的高灵敏度。稳定同位素标记的肝素与来源于动物组织的肝素难以区分，是研究肝素与蛋白质相互作用的新试剂。
We report the first chemoenzymatic synthesis of the stable isotope-enriched heparin from a uniformly labeled [13C,15N]N-acetylheparosan (-GlcA(1,4)GlcNAc-) prepared from E. coli K5. Glycosaminoglycan (GAG) precursors and heparin were formed from N-acetylheparosan by the following steps: chemical N-deacetylation and N-sulfonation leading to N-sulfoheparosan (-GlcA(1,4)GlcNS-); enzyme-catalyzed C5-epimerization and 2-O-sulfonation leading to undersulfated heparin (-IdoA2S(1,4)GlcNS-); enzymatic 6-O-sulfonation leading to the heparin backbone (-IdoA2S(1,4)GlcNS6S-); and selective enzymatic 3-O-sulfonation leading to the anticoagulant heparin, containing the GlcNS6S3S residue. Heteronuclear, multidimensional nuclear magnetic resonance spectroscopy was employed to analyze the chemical composition and solution structure of [13C,15N]N-acetylheparosan, precursors, and heparin. Isotopic enrichment was found to provide well-resolved 13C spectra with the high sensitivity required for conformational studies of these biomolecules. Stable isotope-labeled heparin was indistinguishable from heparin derived from animal tissues and is a novel reagent for studying the interaction of heparin with proteins.
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影响因子：
5.6
作者：
Almond, A;DeAngelis, PL;Blundell, CD
通讯作者：
Blundell, CD
影响因子：
4.1
作者：
Guerrini, Marco;Guglieri, Sara;Mourier, Pierre
通讯作者：
Mourier, Pierre
影响因子：
4.4
作者：
JORGENSEN, WL;CHANDRASEKHAR, J;KLEIN, ML
通讯作者：
KLEIN, ML
DOI：
10.1016/0263-7855(89)80002-5
发表时间：
1989-12-01
期刊：
JOURNAL OF MOLECULAR GRAPHICS
影响因子：
--
作者：
FORSTER, M;JONES, C;MULLOY, B
通讯作者：
MULLOY, B
影响因子：
4.8
作者：
Kuberan, B;Beeler, DL;Rosenberg, RD
通讯作者：
Rosenberg, RD