A guy with a machine: Using NMR to study biological macromolecules

Alexandra A Taylor

doi:10.1074/jbc.cl117.001138

Abstract

A guy with a machine: Using NMR to study biological macromolecules

Highlights

Richards and Moon took advantage of the fact that DPG contains two phosphorus atoms that could be detected using 31P NMR spectroscopy and that the phosphorus signal would be different based on whether or not DPG was bound to hemoglobin, providing an indirect readout of pH. They used this nondestructive technique to peer inside cells and other complex samples, hemolysates, red cell suspensions, and whole blood
The researchers compared the chemical shifts—the relative resonant frequencies of the nuclei— of the phosphorus inside the cells with that in the blood serum to determine the pH inside the red blood cells. They found that 31P NMR is an effective way to determine the pH inside the red blood cell and levels of significant cell metabolites, including ATP, inositol hexaphosphate, and more
Before NMR, researchers had to homogenize cells to get at the intracellular contents, in the process breaking up unique compartments known to have drastically different pH values

Summary

Introduction

Richards and Moon took advantage of the fact that DPG contains two phosphorus atoms that could be detected using 31P NMR spectroscopy and that the phosphorus signal would be different based on whether or not DPG was bound to hemoglobin, providing an indirect readout of pH. They used this nondestructive technique to peer inside cells and other complex samples, hemolysates, red cell suspensions, and whole blood. The researchers compared the chemical shifts—the relative resonant frequencies of the nuclei— of the phosphorus inside the cells with that in the blood serum to determine the pH inside the red blood cells.

Results

Conclusion