Investigation of enzymatic C-P bond formation using multiple quantum HCP nuclear magnetic resonance spectroscopy
Copyright © 2015 John Wiley & Sons, Ltd.
Veröffentlicht in: | Magnetic resonance in chemistry : MRC. - 1985. - 53(2015), 4 vom: 14. Apr., Seite 267-72 |
---|---|
1. Verfasser: | |
Weitere Verfasser: | , , |
Format: | Online-Aufsatz |
Sprache: | English |
Veröffentlicht: |
2015
|
Zugriff auf das übergeordnete Werk: | Magnetic resonance in chemistry : MRC |
Schlagworte: | Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. 13C 1H 31P NMR cobalamin enzymatic reaction mechanisms mehr... |
Zusammenfassung: | Copyright © 2015 John Wiley & Sons, Ltd. 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 (13) C-(31) P multiple quantum (1) H-(13) C-(31) P (HCP) experiment in (1) H-(31) P two-dimensional mode directly on a PhpK-catalyzed reaction mixture using (13) CH3 -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 (13) CH3 and (31) P. Related nuclear magnetic resonance experiments based upon these principles may be designed for the study of enzymatic and/or synthetic chemical reaction mechanisms |
---|---|
Beschreibung: | Date Completed 04.12.2015 Date Revised 13.11.2018 published: Print-Electronic Citation Status MEDLINE |
ISSN: | 1097-458X |
DOI: | 10.1002/mrc.4190 |