Sensitive detection of isoglobo and globo series tetraglycosylceramides in human thymus by ion trap mass spectrometry.
Glycosphingolipids serve as ligands for receptors involved in signal transduction and immune recognition, as exemplified by isoglobotrihexosylceramide, an antigenic ligand for T cell receptors. Mechanistic studies on the regulation of isoglobotrihexosylceramide require biochemical measurement of its lysosomal precursor, isoglobotetraglycosylceramide. It remains a challenge to distinguish between complex tetraglycosylceramide glycosphingolipid isomers with the same sugar components but diverse internal linkages. Here we established a simple and sensitive method to separate globo- and isoglobotetraglycosylceramide by MS5 ion trap mass spectrometry, and report the identification of isoglobotetraglycosylceramide in a CHO cell line transfected by iGb3 synthase, as well as in human thymus.
Li
Yunsen
Y
Department of Melanoma Medical Oncology, The University of Texas M.D. Anderson Cancer Center, Houston, TX 77054, USA.
Teneberg
Susann
S
Thapa
Prakash
P
Bendelac
Albert
A
Levery
Steven B
SB
Zhou
Dapeng
D
D000818:Animals; D016466:CHO Cells; D002675:Child, Preschool; D006224:Cricetinae; D003412:Cricetulus; D005700:Galactosyltransferases / Q000235:genetics / Q000378:metabolism; D005915:Globosides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D018629:Spectrometry, Mass, Secondary Ion; D013950:Thymus Gland / Q000737:chemistry* / Q000378:metabolism; D014162:Transfection
D005915:Globosides; C024032:globotetraosylceramide; C004642:cytolipin R; D005700:Galactosyltransferases; C413211:UDP-galactose beta-D-galactosyl-1,4-glycosylceramide alpha-1,3-galactosyltransferase