Sulfatides isolation

The sulfatide isolated was soluble in chloroform-methanol (2—1) and failed to partition into an aqueous phase. Further identification of the sulfolipid was not made. Goldberg (1961) has postulated several pathways which may be involved in cerebroside sulfate synthesis. The initial steps are the activations of sulfate to 3 -phosphoadenosine-5 -phosphosulfate, i.e., PAPS, as described by Bandurski et al. (1956, 1958), Wilson and Bandurski (1956), and Robbins and Lipmann (1956, 1958a, b). 

Of importance the sulfatides can be further distinguished by a more sophisticated procedure. This would entail intraperitoneal injection of 35S04 into small animals. A quite rapid incorporation of label into brain lipids, for example, will provide support for the presence of a sulfatide. The isolated sulfatide-containing fraction is subjected to thin-layer chromatography, and the lanes are sectioned and assayed for radioactivity. Location of labeled material migrating at the same Rf value of standards (nonlabeled) would be... 

The existence of sulfite oxidase in plants snch as Arabidopsis thaliana (wall or thule cress), tobacco, pea, spinach, barley, carrot, and poplar trees has now been confirmed, and the enzyme from A. thaliana has been structurally characterized. The homodimeric, peroxisomal enzyme lacks the heme domain observed in animal sulfite oxidases, making it the simplest Mo-MPT enzyme yet isolated from eukarya. The animal and plant sulfite oxidases have an overall 47% sequence identity. The detoxification of sulfite, produced in the assimilation of sulfate into sulfur-containing amino acids and membrane components (sulfatides) and from environmental sources (acid rain), is also the principal role of the plant enzymes. 

For the position of the sulfate group, Nakayama96 proposed the primary hydroxyl group of the D-galactopyranosyl group, because of the fact that the sulfatide cannot be tritylated.97 However, the results of methylation experiments disproved this structure. Stoffyn and Stoffyn98 and Yamakawa and coworkers99 isolated 2,4,6-tri-O-methyl-D-... 

Until about ten years ago, this N-(2-hydroxytetracosanoyl)-0-(3-0-sulfato-D-galactopyranosyl)-Clg-sphingosine was the only natural sulfatide known. It can be isolated from the central-nervous material of mammals. 

In the case of acidic glycolipids the relative proton affinity of chemicals can shift the balance for negative ionisation in favor of co-eluted compounds. Pre-analytical separation under acidic conditions serves also to reduce as much as possible the dispersion in the MS spectrum of the metabolite into multiple m/ z representing the various adducts of counterions Na, K, NH4, organic amines, ... which improves sensitivity of the test. Sulfatides are lost during the partition between the hexane and the methanol/water phase. The analysis of sulfatides involves the isolation of the glycosphingolipid fraction and the subsequent separation of sulfatides from neutral lipids by chromatography on DEAE-sephadex or DEAE-cellulose column (the variety of methods are referenced in the website CyberLipid (http //www.cyberlipid.org/). 

Mayers and Haines (1967) have isolated a sulfolipid, 1,14-docosyl disulfate, from the phytoflagellate, Ochromonas danica. Acid hydrolysis of the sulfatide yielded a diol. The structures of the sulfatide (lipoid sulfate ester) and the diol were determined by infrared, NMR, and mass spectroscopy. The infrared spectrum of the sulfatide diol had bands indicative of a long-chain diol 1060cm (primary alcohol CO), 1120 (secondary alcohol CO), and 718 cm (long-chain aliphatic CH2rocking). 

Comparative histochemical and chemical analyses have shown that the accumulated metachromatic material is identical with the sulfatide which has been isolated from brain and other organs of ML (Jatzkewitz 1958, 1960 b Austin 1958, 1959 a, b, 1960 Hagberg et al. 1960) (see below). 

The occurrence of sulfatides in brain was already suspected by Thudichum (1884,1901). Koch (1910) isolated from brain a sulfur-containing Upid which, in... 

Until recently, therefore, the question remained unsettled whether sulfatides contained phosphorus or not. Lees et al. (1959) were able to isolate from brain white matter a sulfatide A in cristalline form, and a sulfatide B which was associated with phospholipids. Similarly Bakke and Cornatzer (1961), and Davison and Gregson (1962) found in normal human brains sulfatides with and without phosphorus upon column chromatographic separation, and the latter authors considered the phosphorus-containing component to result from mere association of sulfatides with phospholipids. In contrast. Green and Robinson (1960), upon paper-chromatography of brain lipid extracts, observed one spot only corresponding to the authentic cerebron sulfate. Today it is generally accepted that sulfatides do not contain phosphorus. 

In 1963, Mehl and Jatzkewitz succeeded in isolating fractions with cerebroside-sulfatase activity from pig kidneys, and Bleszynski and Dzialoszynski (1965) purified soluble arylsulfatases from ox brain. A study by Mehl and Jatzkewitz (1965) on the significance of a lack of arylsulfatase in ML as reported by Austin (1963 a), and Austin et al. (1964 a, b), with 2-hydroxy-5-nitrophenyl-sulfate as substrate, resulted in the finding of two fractions with arylsulfatase activity in normal kidneys. While in ML the smaller of both fractions was present in normal concentration, the second component, and the predominant one in normals, was below the limits (0.005 o. d. units Mehl and Jatzkewitz 1965) of the method in ML. It seems that diminution or lack of this heat-labile fraction, which corresponds to the arylsulfatase A of Austin, is typical for ML, and according to Mehl and Jatzkewitz (1965), supports the assumption of a block in the degradation of ML between sulfatides and cerebrosides (see fig. 3). 

HIV-1 gpl20 V3loop (LAI isolate) CTRPNNN7 RKSIRIQRGPGRAFVTIGKIGNMR QAHC GalCer sulfatide Gb3 GM3 SM... 

HIV-1 gpl20 V3 loop (NDK isolate) CTRPYKVrttQRTSIGLRQSLYTITGKKKKtTGVIGQAHC GalCer sulfatide Gbs GM3 SM... 

HIV-2 gplOB V3 loop (ROD Isolate) CKRPGNKTV QIMLMSGHVFHSHyQPINKRPRlQAWC GalCer sulfatide CbjGM3... 

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