A novel disialoganglioside in rat spleen lymphocytes.

K Nohara,M Suzuki,F Inagaki,T Sano,K Kaya

doi:10.1016/s0021-9258(18)42136-9

Abstract

A novel ganglioside has been identified as the predominant disialoganglioside of the lymphocytes prepared from rat spleen. The ganglioside was isolated from rat spleen and characterized by compositional analysis, methylation analysis, sialidase hydrolysis, proton NMR spectroscopy, and negative ion fast atom bombardment mass spectrometry. The structure was determined as follows. [formula: see text] This ganglioside is a unique derivative of N-acetyllactosaminyl-GM1. The three monosialogangliosides containing N-acetyllactosaminyl-GM1 structure, which had been originally isolated from rat spleen (Nohara, K., Suzuki, M., Inagaki, F., Ito, H., and Kaya, K. (1990) J. Biol. Chem. 265, 14335-14339), were also found in the lymphocytes and were hardly detected in the spleen remnant tissue depleted of single cells. On the other hand, GD1c(NeuGc,NeuGc) (IV3(NeuGc alpha 2-8NeuGc)-Gg4Cer), the overwhelmingly predominant ganglioside of rat thymocytes (Nohara, K., Suzuki, M., Inagaki, F., and Kaya, K. (1991) J. Biochem. (Tokyo) 110, 274-278), was demonstrated to be only a minor component of the gangliosides of the spleen lymphocytes. These results suggested that GD1c is characteristic for the immature T lineage lymphoid cells and the gangliosides having lactosaminyl-GM1 structure are specific for other populations of the lymphocytes in rat.

Highlights

From the $Environmental Health Sciences Division, an1d1Environmental Chemistry Division, The National Institute for Environmental Studies,Onogawa, Tsukuba, Ibaraki305 and the Departments of §Membrane Biochemistry and 7Molecular Physiology, The Tokyo Metropolitan Instituteof Medical Science, Bunkyo-ku, Tokyo113, Japan
A novel ganglioside has been identified as the pre- is a unique GMlb-derivedldisialoganglioside, GD,(NeuGc, dominant disialoganglioside of the lymphocytes pre- NeuGc) [8]
Theganglioside was isolated In thepresent study,we have characterized a novel gangliofrom rat spleen and characterized by compositional side as themajor disialogangliosideof rat spleen lymphocytes

Summary

RESULTS

(9:l) and(B) 1-propanol/l.5 N NH,OH [64] changing the B content The lymphocytes were prepared from rat spleen using Fifrom 10 to 50% for 400 min a t a flow rate of0.6 ml/min. The gangliosides were obtained from the cells and compared with thofsreom the spleen remnant tissue which were depleted of single cells (Fig. 1). All the gangliosides of the lymphocytes were fractionated into. -GMl mono- and disialogangliosides by DEAE-Sephadex column chromatography (Fig. 1, lunes 1 and 2). Cytes were hardly detected(Fig. 1, lune 4 ) .Among the disialogangliosides from the lymphocytes, the ganglioside with RF value between that of G o l h and G T I h was most abundant. The disialoganglioside wasdetected at a much lower concentration in the remnant tissue than in the lymphocytes Form/methanol/0.2% CaCI2 (60409) in a , chloroform/methanol/2.5 N NH,OH (60:40:9) in b, anld-propanol/28%.

GOlb GTlb

Position of

GanLgylmiopSsihpdoleecseyontfeRs at VI v

Findings

GM GDla