Combining solid-state and solution-state 31P NMR to study in vivo phosphorus metabolism.
Otherwise unavailable information concerning the distribution of phosphorylated compounds in biological systems is obtained by a combined solid-state/solution-state NMR approach, illustrated here for oocytes from Rana pipiens. General methodology is developed, and further extensions are proposed. The following conclusions pertain to the specific system under examination. (i) Nucleoside phosphates can be observed by magic-angle sample spinning of the lyophilized material. (ii) The solid-state NMR technique of dipolar decoupling provides no additional resolution of the phospholipid and phosphoprotein components of the yolk. However, cellular death produces sufficient pH changes to cause the phospholipid and protein phosphate peaks to become resolvable. The concentration of nucleoside phosphates also decreases. (iii) The phospholipid and phosphoprotein components are shown by computer simulation to be present in a ratio of 40:60, respectively. (iv) The amounts of inorganic phosphate, nucleoside phosphates, and sugar phosphates are determined by solution-state NMR observation of the perchloric acid extract of the oocytes.
D000818:Animals; D002470:Cell Survival; D004530:Egg Yolk / Q000032:analysis; D005260:Female; D005612:Freeze Drying; D009682:Magnetic Resonance Spectroscopy; D009865:Oocytes / Q000378:metabolism; D010758:Phosphorus / Q000378:metabolism*; D011894:Rana pipiens; D012995:Solubility
D010758:Phosphorus