Mono-sulfated globotetraosylceramide from human kidney.
A novel sulfated glycosphingolipid that belongs to "globo-series" was isolated from human kidney. This lipid was purified from a pooled kidney preparation by chloroform-methanol extraction, mild alkaline treatment, DEAE-Sephadex and silicic acid column chromatographies, and preparative TLC. The structure and the properties were studied by IR spectroscopy, proton NMR spectroscopy, negative secondary ion-mass spectrometry, solvolysis, periodate oxidation, compositional and methylation analyses, monoclonal antibodies, and a sulfatide-binding protein. From the results of the above analyses, the structure of this glycolipid was proposed to be HSO3-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1-1ceramide. This sulfated lipid reacted with a monoclonal anti-SSEA-3 (stage-specific embryonic antigen-3) (MC-631) (Kannagi, R., Cochran, N.A., Ishigami, F., Hakomori, S., Andrews, P.W., Knowles, B.B., & Solter, D. (1983) EMBO J. 2, 2355-2361), whose epitope is R-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-R', on TLC and solid-phase radioimmunoassay. This lipid also bound to the 125I-labeled sulfatide-binding protein, thrombospondin. The yield of this sulfated glycolipid was 34 pmol/g of tissue, which was about 0.028, 0.16, and 18 mol% of galactosyl- and lactosylceramide sulfates, and globopentosylceramide sulfate (Nagai, K.-i., Roberts, D.D., Toida, T., Matsumoto, H., Kushi, Y., Handa, S., & Ishizuka, I. (1989) J. Biol. Chem. 264, in press), respectively, in human kidney.
Nagai
K
K
Department of Biochemistry, Teikyo University School of Medicine, Tokyo.
Roberts
D D
DD
Toida
T
T
Matsumoto
H
H
Kushi
Y
Y
Handa
S
S
Ishizuka
I
I
D000818:Animals; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins / Q000378:metabolism; D002855:Chromatography, Thin Layer; D008401:Gas Chromatography-Mass Spectrometry; D005915:Globosides / Q000302:isolation & purification*; D006028:Glycosphingolipids / Q000302:isolation & purification*; D006801:Humans; D007668:Kidney / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins / Q000378:metabolism; D008745:Methylation; D010084:Oxidation-Reduction; D010504:Periodic Acid; D011863:Radioimmunoassay; D051381:Rats; D013055:Spectrophotometry, Infrared; D013433:Sulfoglycosphingolipids / Q000302:isolation & purification; D019699:Thrombospondins
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D005915:Globosides; D006028:Glycosphingolipids; D008562:Membrane Glycoproteins; D013433:Sulfoglycosphingolipids; D019699:Thrombospondins; D010504:Periodic Acid; C024032:globotetraosylceramide; C009288:metaperiodate