31P-NMR Spectroscopy of Glucose-6-phosphate, Glucose-1-phosphate and NADH

The addition of phosphate groups (-OPO32-) into carbohydrates is one of many central biochemical processes to regulate the sugar household within the human body. The sugar household contains the body’s intake of sugars and starches (carbohydrates) to provide the necessary energy to function properly. Common glycosyl phosphates such as glucose-1-phosphate, glucose-6-phosphate, and NADH were analyzed by magnetic resonance (NMR) spectroscopy at 600 MHz to determine multiple types of coupling constants (1H-13C, 1H-31P, and 13C-31P) and to draw conclusions about the three-dimensional position of the phosphate group in each case. Through total correlation spectroscopy (TOCSY), correlated spectroscopy (COSY), heteronuclear multiple bond correlation (HMBC), J-resolved heteronuclear multiple bond correlation (J-HMBC), and heteronuclear single-quantum coherence multiple bond correlation (HSQMBC), the structure of each glycosyl phosphate was further studied to determine coupling constants from the phosphate group to the neighboring carbon and hydrogen atoms. In the analyzed data, it was observed that the phosphate group has a preferred conformation relative to the sugar ring. The so-established NMR experiments were subsequently applied to NADH and allowed us to complete the three-dimensional structure assignment