Application of Two-Dimensional Total Correlation Spectroscopy for Structure Determination of Individual Inositol Phosphates in a Mixture
✍ Scribed by Kermit Johnson; Laura G. Barrientos; Long Le; Pushpalatha P.N. Murthy
- Publisher
- Elsevier Science
- Year
- 1995
- Tongue
- English
- Weight
- 204 KB
- Volume
- 231
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
and references therein). Establishing the specificity of The discovery of the second messenger role of myoinositol phosphate-metabolizing enzymes has always inositol(1,4,5)trisphosphate has triggered tremenbeen a challenge because of the need to separate and dous interest in investigating the structures and meidentify the structures of individual inositol phostabolism of inositol phosphates. The structures of phates in a mixture containing stereo-and regioisothese compounds are established by first purifying mers. This problem is particularly acute when investithe compounds of interest and then identifying the gating the specificity of phytases, a group of enzymes structures through chemical degradation or NMR that hydrolyze phytic acid to a series of lower phosanalysis. In this paper we describe a method that phoric acid esters of myo-inositol and, in some cases, allows simultaneous determination, without separafinally to myo-inositol itself (9-11).
tion, of the structures of multiple inositol phos-
Structures of inositol phosphates have been estab-
phates in a mixture. The use of two-dimensional total lished by first purifying the compound, which often recorrelation spectroscopy (2D TOCSY) experiments quires employing more than one technique (HVE, together with subspectra extracted from 2D TOCSY HPLC, and/or column chromatography), and then subdata allowed us to determine, in 3 to 4 h, the strucjecting the purified compound to elaborate chemical tures of five compounds in a mixture, without prior and enzymatic reactions and identifying the products separation.