Galactosylceramide containing omega-amino-fatty acids: preparation, characterization, and sulfotransferase acceptor.
For preparation of an affinity ligand, an N-fatty acyl moiety of galactosylceramide (GalCer) was chemically replaced with omega-amino-fatty acid including amino-n-hexanoic acid or amino-n-dodecanoic acid to obtain omega-aminoGalCer. For the synthesis of the compound, galactosylsphingosine (GalSph) was coupled with N-trifluoroacetyl omega-amino-fatty acid which was prepared by a reaction with S-ethyltrifluorothioacetate. After removal of the N-trifluoroacetyl group in a mild alkaline solution, in which an N-fatty acyl group was retained, aminoGalCer composed of an N-hexanoyl or an N-dodecanoyl group was obtained with an overall yield of 90%. Their chemical structures were confirmed by proton nuclear magnetic resonance and fast atom bombardment-mass spectrometries. These aminoGalCers and GalSph as well as immobilized aminoGalCer were sulfated by a glycolipid sulfotransferase from rat kidney. Furthermore, immobilized aminoGalCer on gel matrix was used for affinity chromatography of the sulfotransferase, resulting in an excellent increase in the purification (14,000-fold) with a recovery rate of 40%.
Kamio
K
K
Biochemistry Laboratory, Hokkaido University School of Medicine, Sapporo, Japan.
Gasa
S
S
Makita
A
A
D015119:Aminocaproic Acid / Q000737:chemistry; D000818:Animals; D005699:Galactosylceramides / Q000737:chemistry* / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology*; D007850:Lauric Acids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D051381:Rats; D016339:Spectrometry, Mass, Fast Atom Bombardment; D015238:Sulfotransferases / Q000302:isolation & purification / Q000378:metabolism*
D005699:Galactosylceramides; D007850:Lauric Acids; C046049:12-aminododecanoic acid; D015238:Sulfotransferases; D015119:Aminocaproic Acid