Investigation of enzymatic C-P bond formation using multiple quantum HCP nuclear magnetic resonance spectroscopy
Hu, Kaifeng1,2; Werner, Williard J.3; Allen, Kylie D.3,4; Wang, Susan C.3; Hu,KF (reprint author),Chinese Acad Sci,Kunming Inst Bot,State Key Lab Phytochem & Plant Resources West Ch,Kunming 650201,Yunnan,Peoples R China.; kaifenghu@mail.kib.ac.cn; susan_wang@wsu.edu
2015-04-01
发表期刊MAGNETIC RESONANCE IN CHEMISTRY
卷号53期号:4页码:267-272
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.
关键词Nmr H-1 C-13 P-31 Phosphinates Methylation Enzymatic Reaction Mechanisms Cobalamin Radical S-adenosyl- L-methionine
学科领域Chemistry, Multidisciplinary ; Chemistry, Physical ; Spectroscopy
DOI10.1002/mrc.4190
收录类别SCI
语种英语
WOS记录号WOS:000351636200004
引用统计
文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/20585
专题植物化学与西部植物资源持续利用国家重点实验室
通讯作者Hu,KF (reprint author),Chinese Acad Sci,Kunming Inst Bot,State Key Lab Phytochem & Plant Resources West Ch,Kunming 650201,Yunnan,Peoples R China.; kaifenghu@mail.kib.ac.cn; susan_wang@wsu.edu
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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Hu, Kaifeng,Werner, Williard J.,Allen, Kylie D.,et al. Investigation of enzymatic C-P bond formation using multiple quantum HCP nuclear magnetic resonance spectroscopy[J]. MAGNETIC RESONANCE IN CHEMISTRY,2015,53(4):267-272.
APA Hu, Kaifeng.,Werner, Williard J..,Allen, Kylie D..,Wang, Susan C..,Hu,KF .,...&susan_wang@wsu.edu.(2015).Investigation of enzymatic C-P bond formation using multiple quantum HCP nuclear magnetic resonance spectroscopy.MAGNETIC RESONANCE IN CHEMISTRY,53(4),267-272.
MLA Hu, Kaifeng,et al."Investigation of enzymatic C-P bond formation using multiple quantum HCP nuclear magnetic resonance spectroscopy".MAGNETIC RESONANCE IN CHEMISTRY 53.4(2015):267-272.
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