Determination of glucan phosphorylation using heteronuclear 1H, 13C double and 1H, 13C, 31P triple-resonance NMR spectra

Magn Reson Chem. 2013 Oct;51(10):655-61. doi: 10.1002/mrc.3996. Epub 2013 Aug 4.

Abstract

Phosphorylation and dephosphorylation of starch and glycogen are important for their physicochemical properties and also their physiological functions. It is therefore desirable to reliably determine the phosphorylation sites. Heteronuclear multidimensional NMR-spectroscopy is in principle a straightforward analytical approach even for complex carbohydrate molecules. With heterogeneous samples from natural sources, however, the task becomes more difficult because a full assignment of the resonances of the carbohydrates is impossible to obtain. Here, we show that the combination of heteronuclear (1) H,(13) C and (1) H,(13) C,(31) P techniques and information derived from spectra of a set of reference compounds can lead to an unambiguous determination of the phosphorylation sites even in heterogeneous samples.

Keywords: heteronuclear NMR; phosphorylation; starch; triple resonance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Isotopes
  • Glucans / chemical synthesis*
  • Glucans / chemistry
  • Magnetic Resonance Spectroscopy / standards
  • Phosphorus Isotopes
  • Phosphorylation
  • Protons
  • Reference Standards

Substances

  • Carbon Isotopes
  • Glucans
  • Phosphorus Isotopes
  • Protons