A1H/13C inverse 2D method for the analysis of the polyamines putrescine, spermidine and spermine in cell extracts and biofluids

Abstract

The polyamines putrescine, spermidine and spermine are involved in the regulation of various metabolic processes. It is therefore desirable to detect and quantify the polyamines with NMR. We present the proton and carbon assignments for all polyamine signals obtained from PCA extracts of F98 glioma cells with high resolution using a semi‐selective HSQC 2D‐experiment. The biosynthesis of the polyamines in cell culture was examined using the labeled substrates [U‐13 C]glucose and [U‐13 C]glutamate. In such studies the high resolution of the semi‐selective HSQC experiment at very high magnetic fields (14–19 T) allows the analysis of carbon‐carbon couplings, and isotopomer patterns. The different effects of osmotic stress on the concentrations of polyamines and amino acids are also reported. © 1998 John Wiley & Sons, Ltd.

Non‐destructive quantitation of spermine in human prostate tissue samples using HRMAS 1H NMR spectroscopy at 9.4 T

“…The signal assignments are based on both the literature [5–8] and our own measurements of authentic compounds of spermine, spermidine, and putrescine using the same experimental conditions as those used for tissue analysis (data not shown). Although other polyamines (putrescine and spermidine) would also have resonances in the 3.09–3.13 ppm region, our results confirmed that spermine is the predominant polyamine in prostate as previously reported [11–13].…”

900MHz 1H-/13C-NMR analysis of 2-hydroxyglutarate and other brain metabolites in human brain tumor tissue extracts

“…Although the 1 CH 2 and 2 CH 2 peaks from the putrescine moieties of spermine and spermidine were not distinguishable from each other, the 3 CH 2 and 4 CH 2 peaks from spermidine were separately identifiable. The positions of the HSQC cross peak of both spermine and spermidine showed excellent agreement with previously published data after adjusting chemical shift reference [ 34 ]. Free putrescine might be present, but even in 1 H- 13 C HSQC the peak positions are almost identical to those of spermidine so that it could not be separately observed [ 34 ].…”

“…The positions of the HSQC cross peak of both spermine and spermidine showed excellent agreement with previously published data after adjusting chemical shift reference [ 34 ]. Free putrescine might be present, but even in 1 H- 13 C HSQC the peak positions are almost identical to those of spermidine so that it could not be separately observed [ 34 ]. The outermost 3 CH 2 peaks (the 3 CH 2 peaks from two 3-aminopropyl moieties in spermine and one in spermidine) appeared at almost identical position to the one from β CH of phenylalanine in HSQC.…”

In vivo dynamic turnover of cerebral 13C isotopomers from [U–13C]glucose

“…The isotopomer patterns observed in vivo are consistent with the known 13 C-13 C homonuclear J couplings previously detected in vitro [5]. At early time points, the predominant isotopomer at glutamate C4 at 34.00 ppm is Glu4D45 contributed by [4,[5][6][7][8][9][10][11][12][13] C]glutamate which is formed by the condensation of [1, 2-13 C]acetylCoA with unlabeled oxaloacetate during the first turn of the TCA cycle. Then Glu4Q gradually dominates the isotopomer pattern of glutamate C4 as the labeling of the carbonyl carbon of oxaloacetate increases over time.…”