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题名: Investigation of enzymatic C-P bond formation using multiple quantum HCP nuclear magnetic resonance spectroscopy
作者: Kaifeng Hu(胡凯锋)1, 2; Werner, Williard J.3; Allen, Kylie D.3, 4; Wang, Susan C.3
刊名: MAGNETIC RESONANCE IN CHEMISTRY
关键词: NMR ; H-1 ; C-13 ; P-31 ; phosphinates ; methylation ; enzymatic reaction mechanisms ; cobalamin ; radical S-adenosyl- l-methionine
英文摘要: The biochemical mechanism for the formation of the C-P-C bond sequence found in l-phosphinothricin, a natural product with antibiotic and herbicidal activity, remains unclear. To obtain further insight into the catalytic mechanism of PhpK, the P-methyltransferase responsible for the formation of the second C-P bond in l-phosphinothricin, we utilized a combination of stable isotopes and two-dimensional nuclear magnetic resonance spectroscopy. Exploiting the newly emerged Bruker QCI probe (Bruker Corp.), we specifically designed and ran a C-13-P-31 multiple quantum H-1-C-13-P-31 (HCP) experiment in H-1-P-31 two-dimensional mode directly on a PhpK-catalyzed reaction mixture using (CH3)-C-13-labeled methylcobalamin as the methyl group donor. This method is particularly advantageous because minimal sample purification is needed to maximize product visualization. The observed 3:1:1:3 multiplet specifically and unequivocally illustrates direct bond formation between (CH3)-C-13 and P-31. Related nuclear magnetic resonance experiments based upon these principles may be designed for the study of enzymatic and/or synthetic chemical reaction mechanisms. Copyright (c) 2015 John Wiley & Sons, Ltd.
出版日期: 2015-04-01
卷号: 53, 期号:4, 页码:267-272
DOI标识: 10.1002/mrc.4190
语种: 英语
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内容类型: 期刊论文
URI标识: http://ir.kib.ac.cn/handle/151853/20585
Appears in Collections:植物化学与西部植物资源持续利用国家重点实验室_期刊论文

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作者单位: 1.Chinese Acad Sci, Kunming Inst Bot, State Key Lab Phytochem & Plant Resources West Ch, Kunming 650201, Yunnan, Peoples R China
2.Univ Wisconsin, Dept Biochem, Natl Magnet Resonance Facil Madison, Madison, WI 53706 USA
3.Washington State Univ, Sch Mol Biosci, Coll Vet Med, Pullman, WA 99164 USA
4.Virginia Polytech Inst & State Univ, Dept Biochem, Blacksburg, VA 24061 USA
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