Ganglioside mixtures

V. B. Ivleva, Y. N. Elkin, B. A. Budnik, S. C. Moyer, P. B. O Connor, and C. E. Costello, Coupling thin-layer chromatography with vibrational cooling matrix-assisted laser desorption/ ionization Fourier transform mass spectrometry for the analysis of ganglioside mixtures, Anal Chem., 76 (2004) 6484-6491. 

Recently, diethylaminoethyl (DEAE) silica gel was shown to be an improvement over typical cellulosic-matrices resins for the separation of acidic and neutral lipids from complex ganglioside mixtures. The specific advantages claimed were ... 

The mechanically fragmented brain or spinal cord was dehydrated with acetone, as in the isolation of cerebrosides.7 Gangliosides are present in the water-soluble fraction they were recognized by color reactions (for example, Bial s orcinol reaction) based on the presence of sialic acid.159 The ganglioside mixture may be purified by partition... 

Fig. 4.5.10. Elution profile of total gangliosides from beef brain on DEAE-silica gel (acetate form) [93]. Bed dimensions 60x2 cm. Sample applied, 40 mg ganglioside mixture. Flow-rate 1.5 ml/min. Eluents methanol, 0.2 M ammonium acetate in methanol and 0.5 M ammonium acetate in methanol gradient elution. Fractions of 15 ml were collected. Resorcinol assay method for detection of sialic acid. Mono- to tetra- denote the numbers of sialic acid residues in the ganglioside fractions. The gangliosides were isolated after removal of ammonium acetate by dialysis against cold water.

TLC HPTIjC plates Silica—aluminum T JC CHa3/MeOH/0.2% CaClj (55 45 10, THAP Matrix mixtures MALDI/FT-MS Ganglioside mixtures [39] ... 

Total ganglioside mixtures (Porcine Brain) (Avanti Polar Lipids, Inc., Alabaster, AL, USA). 

The protein kinase activity which was stimulated in rats by cholera entero-toxin was blocked by the addition of a ganglioside mixture. The same gangliosides were, however, not directly involved in the binding of gonadotrophin to ovarian cell membranes. 

Another major constituent of the brain ganglioside mixture is ganglioside C3(=C3=Ggntiii)- This molecule has the basic structure of B3, but has one additional neuraminic acid residue linked by a 28-bond to the N-acetyl-neuraminic... 

The other gangliosides listed in the table are minor components of the total ganglioside mixture. They differ in the size of the hydrophilic carbohydrate part and in the number of N-acetyl-neuraminic acid molecules. The different chromatographic behaviour, particularly in thin-layer chromatography allows their easy detection. A gangUoside with a simple structure has been isolated from horse erythrocytes (Yamakawa 1951 Klenk et al. 1952). It has been named haematoside and its exact structure determined. N-acetyl-neuraminic acid is substituted by N-glycolyl-neuraminic acid ... 

Fig. 1 Protein concentration dependence of X-ray scattering curve I q) of albumin/ganglioside mixture suspensions at 25 °C. (a) and (b), albumin/G i ganglioside mixture in 50 mM hepes buffer at pH 7.0 (c) and (d), albumin/Gma ganglioside mixture in 0.01 M citrate buffer at pH 6.7. The ganglioside concentration was 0.5% w/v and the albumin concentration was varied. Two different types of albumins were added (a) and (c), albumin-2-amido-2-deoxy-D-galactose (b) and (d), albumin-N-l-[deoxymaltitol]...

Fig. 2 Distance distribution function p(r) of albumin/ganglioside mixture suspension calculated by the Fourier inversion of the scattering curve I q) in Fig. 1. (a)-(d) as in Fig. 1...

Tay-Sachs ganglioside noimally represents approximately 10% of the total ganglioside mixture. In Tay-Sachs disease where there is a tenfold increase in total cerebral ganglioside content, this species represents 90% of the mixture (Svennerholm, 1962 Klenk, 1963). 

In addition to enabling separate analysis of at least two types of sialic acid there are the additional advantages inherent in GLC assay sensitivity and specificity. As little as 1 fig of lipid-bound sialic acid could be conveniently analyzed and this could be reduced to 0.3/Ltg and perhaps less with suitable controls for background peaks. For routine assays OV-1 has been the column of choice, but the somewhat more polar OV-225 has proved quite useful in confirming peak identification where required. A typical analysis with both columns is shown in Figure 14 the sample was purified ganglioside mixture from ovine adrenal medulla (Price and Yu, 1976), which was found to contain NANA and NONA in a ratio of approximately 3 2, respectively. 

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